the market source for life science...

Gel tank and blotting systemin one from C.B.S. Scientific

page 30

Get custom bar coding on yourdeep-well plates from Brand

page 8

Accelerate protein samplepreparation and analysis fromPall Life Sciences

page 12

Beat genomic DNA contamination back with E.Z.N.A.TM RNA extraction system from Omega Bio-tek

page 36

ISSUE 17 - SPRING 2007The Market Source for Life Science

vwr.com

Shop!Make sure you get all our special

offers - just for lifescientists with

the VWRbioMarke shop or register

for the VWRbioMarke e-newsletter

on [email protected]

Abgene*

Air Liquide

AppliChem

Axygen

BD Biosciences

Binder

Brand

BTX Harvard Apparatus

CBS

Merck Biosciences

Molecular BioProducts

Nalgene

Nunc

Omega Bio-Tek

Operon

Pall Life Sciences

5 PRIME

Sartorius

Spectrum

Thermo Scientific

Whatman

Wheaton

ABgene* products are not availablein Italy, Belgium and the Netherlands

table of contentsCell banking and compound storage with high recovery vials, Wheaton 3

Simulation of natural conditions in CO2 incubators, Binder 4

Human embryonic stem cells: the next step, BD Biosciences 6

Individual bar coding of deep well plates, Brand 8

OptiCell® cell culture system, Nunc 10

Livecell ArrayTM, Nunc 11

Ultimate flexibility for your protein quantification needs, Thermo Scientific 14

High sensitivity DNA and protein analysis with Multiskan® Spectrum, Thermo Scientific 16

The new Krosflo® research II TFF system, Spectrum Labs 18

MIEF-SYS Isoelectric focusing horizontal electrophoresis unit, VWR Collection 20

New! product portfolio, 5 PRIME 22

Speed your throughput with blotting sandwiches, Whatman 23

New insights into DNA contamination , Applichem 24

Traceability of biological sample storage, Air Liquide 26

Tecan compatible tips, Axygen 28

New tip refill system, Axygen 29

The Dual Cool electrophoresis system, CBS 30

D-TubeTM dialysers and D-Tube electroelution kit, Merck Biosciences 32

The OpScanner for all standard microarray slides, Operon 34

Sample preparation for drug testing in hair, Sartorius 35

High quality RNA isolation using E.Z.N.A TM RNA system, Omega 36

Accelerate protein sample preparation and analysis, Pall Life Sciences 12

Control the power - transfer the genes, BTX 39

AbsoluteTM Blue - the advantage is clear, Abgene 40

Contamination control - low retention tips, MBP 42

Make your research efforts more efficient! VWRBioSciences 44

VWR I n t e r n a t i o n a l Issue 17 A p r i l 2 0 0 72

editorialThis is the first edition of the VWRbioMarke Magazine for 2007, where we welcome back ourVWRbioMarke Partners bringing the high quality of innovation and application support that makesthem eligible to join this unique programme crafted for life scientists.

We are very excited this year to include some new names5-Prime – for the Eppendorf 5-Prime reagent portfolio as well as a selection of products from the Gentra product portfolio from QIAGENApplichem – a well etablished manufacturer of a broad range of biochemicals for biochemistry, cell and molecular biologyAnd CBS-market leading Electrophoresis rejoin the programme to being you some stunning newdevelopments in PAGE.

Watch this space for new VWRbioMarke partners as the year progresses, we will be working hardbehind the scenes to bring you even more!

The VWR Collection is going from strength to strength as a force in the marketplace for great valueat competitive prices and in the lifescience area this year there will be some fantastic new products.In this issue we include some information on a new IsoElectric focusing system.

We hope that you find this issue a useful contribution to support your work and to further meet yourneeds check out the companion promotional flyer – the VWRbioMarke Shop that includes a widevariety of offers and savings on the products you use everyday.

Enjoy the spring!The VWRbioMarke Team

EditorVWR International Europe bvbaHaasrode Researchpark Zone 3Geldenaaksebaan 4643001 LeuvenBelgium

CopywritingVWR International Europe bvba

Layout and typesettingMarketing Services VWR

PrintingStork, GermanyNo part of this publication may be reproduced or copied withoutprior permission by writing of VWR International Europe.

Run77.650 copiesPublication date: April 2007

Due to the high sales volume of promoted articles someitems may be temporarily out of stock - VWR Terms andConditions of Sale apply.

There is an increasing need

among biopharmaceutical

companies to utilise an

integrated systems approach

towards sample management in

compound libraries and cell

banks. The number of

compounds generated for

compound libraries and cell

banks continues to expand

yearly, necessitating seamless

integration of storage vials with

compound library robotics

systems in order to track,

transport and process the

materials without compromising

the performance of the robotic

systems. The Wheaton

E-Z Ex-TractionTM vial has been

designed to meet these needs.

VWR I n t e r n a t i o n a l Issue 17 A p r i l 2 0 0 7 3

For more information on these products contact your local VWR sales office,send an e-mail to [email protected] or visit our website www.vwr.com

The Wheaton Science Products patented E-Z Ex-Traction™ vials are made from 33expansion borosilicate glass. This glass is widelyused because it has low leachables / extractables.These vials are designed for the storage of highvalue samples. The bottom of the vial has a conicalshape that allows for more complete recovery ofthe sample material than can be retrieved fromstandard flat bottom sample vials. The conicalbottom allows for the syringe or cannula to alignand remove nearly all of the material contained inthe vial. The Wheaton quality control processensures that the conical point is at the exactcentre of the vial and at a consistent depth toassure that the needle or cannula portion of anautomated sampler will not jam when the needleis inserted into the vial. In addition, the flat,smooth vial base is designed to easilyaccommodate a 2-D barcode.

The patented bottom is manufactured with aconstant depth and shape, critical for automatedsystems easily damaged by irregularities in vialdimensions. The interior walls provide a smoothtransition into the conical well at the bottom ofthe vial, which prevents loss of materials thatotherwise may collect on ledges. The generalprofile of the E-Z Ex-Traction™ vial is designed towork well with commercial automated storageand liquid handling systems. The vial allowssamples to move from storage through topreparation and analysis without needing to betransferred to other containers.

E-Z Ex-Traction™ vials can be supplied in eitherclear or amber glass. Amber vials are used for lightsensitive samples. The vials sizes currentlyavailable are: 2, 4, 8, 10, and 20 ml. Wheaton E-Z Ex-Traction™ vials are available with a screwthread finish. Other vial sizes and serum finishescan be created as part of Wheaton’s Sci-Lutions™offering. Vials can be supplied bar coded, tared,sterilized, critically cleaned or silanised to fitindividual application requirements.

The E-Z Ex-Traction™ vial is advantageous inquality assurance testing for the production ofmonoclonal antibodies as a sampling vial becausethe conical bottom requires less MAb to besampled than corresponding flat bottom vials. Useof the E-Z Ex-Traction™ vial as a sampling vial inaffinity chromatography QC, cat ion exchangechromatography QC, anion exchangechromatography QC and other down streamprocess QC protocols may result in substantialsavings of MAb product as less product will bewasted while performing the required QC testingprotocols.

Manufacturers of MAb products will also find theE-Z Ex-Traction™ vial to be a suitable containerfor single dose packaging if customised with aserum finish. High volume users may requestcustom modifications of E-Z Ex-Traction™ vials tofit them to their particular applications, whetherfor storage of sample materials or for packaging aproduct.

Cell banking and compound storage with high recovery vials

Why use Wheaton’s high recovery vials?

Nature sets the standard

TemperatureTemperature is a decisive factor for growth. Strong cell growth can be achieved only within a verynarrow tolerance range around the optimal temperature.

Standard at BINDER: the VENTAIR™ jacket system - ensures fully homogenous temperature controlof the complete interior. The BINDER Airflow Design and Intelligent Temperature Control for optimaltemperature stability for natural temperature conditions.

CO2 concentrationConventional measurement systems basically measure the CO2 content as a function of humidity.Both humidity and CO2 concentration change when the door is opened. Such measurements can beincorrect in real time. IR measurement technology, which is the professional standard at BINDER,prevents such deviations.

Standard at BINDER: the CO2 infrared measurement system - a single-beam differentialmeasurement with a drift-free FPI sensor.Direct and fast permanent digital CO2 gas measurement. Indispensable for precise andfast adjustment of the required CO2 content. Result: up to 20 times faster CO2 content recoverytimes compared to thermoconductivity measurements.This measurement process does not require time-consuming and imprecise auto-zero calibration.Patented gas mixing head. This actually provides completely homogenous gas distributionwithout the use of fans to prevent turbulence and mixing of germs in the interior.

Humidity

Naturally, high saturated air humidity is important. It prevents cell cultures from drying out and alsokeeps the osmolarity constant in the culture medium.

Standard at BINDER: the Permadry™ systemThe Permadry™ double pan system with defined condensation point is a simple and innovativehumidification principle for maximum humidity values of >95% rH with completely dry chamberwalls at the same time.

BINDER CO2 incubatorssimulate natural conditions,where many perfect detailsmake up the sum total. We also fuse together thedetails by amalgamating thegrowth parameters such asCO2, temperature andhumidity into a finely tunedinteraction that we call“natural simulation.” This is a process that we havedeveloped and patented,unparalleled in our industryand the closest thing tonatural conditions.


Nature always sets the standard, but we

VENTAIR™ jacket system The CO2 infrared measurement system Permadry™ double pan system



come very close

Cell culture

>Standard equipment of the BINDER CB series

• Temperature range of 7 °C (13 °F) above ambient temperature up to 60 °C (140 °F)

• MCS controller for temperature and CO2 concentration

• Standard-compliant hot-air sterilisation at 180 °C (DIN 58947)

• Weldless deep-drawn inner chamber made of stainless steel with integrated shelf support system. Copper models with dismantable shelf mounting system.

• Electronic self-diagnostic system for errors with optical and acoustic alarm, as well asrelay contact for central monitoring

• Tightly closing inner glass door

• RS 422 interface for communication software APT-COM™ DataControlSystem

• BINDER test certificate

Numerous research efforts are being expendedon establishing suitable procedures for the invitro cultivation and expansion of hES cells.Initial culturing techniques originated frommethodology developed for murine ES (mES)cells and included the use of mouse embryonicfibroblast (MEF) feeder layers. While MEFfeeder-based systems have proven to berobust for the long-term culturing of hES cells,they present a number of limitations. Concernsabout contamination of hES cells with animal-derived pathogens from MEF feeder layersnotably make these culture systems unsuitablefor clinical applications.

Embryonic stem cells (ES cells) have

an infinite capacity for self-renewal

and the remarkable ability to

differentiate into virtually all cell

types of the adult body. Human ES

(hES) cells can differentiate into

any cell type represented by the

three germ layers – ectoderm,

mesoderm and endoderm. Because

of their unique combined abilities

of infinite expansion and

pluripotency, hES cells hold great

promise for application in

functional genomics, cell therapy

and regenerative medicine.


Human Embryonic Stem Cells:The next stepLong-term propagation of human embryonic stem (hES) cells: feeder-free, streamlined and reproducible.

Figure 1. Representative phase contrast images of H9 hES cells grown on a) MEF feeder layers and b) BD BioCoatTM

MatrigelTM Matrix Plates for ES Cell Culture. hES cells appear to spread out more and form larger, monolayer-like colonieson the BD MatrigelTM Matrix surface, compared to those cultured on MEF feeder layers; c) embryoid bodies (EBs) formedwithin 48-72 hours upon transferring H9 cells cultured on BD BioCoatTM MatrigelTM Matrix Plates for ES Cell Culture for 10 passages onto a low-attachment surface* and culturing them in media supplemented with FBS and without bFGF.

* low-attachment surface available through BD Biosciences’ Custom Coating Service. Please contact [email protected] for more information.

When choosing BD BioCoat™ Matrigel™ Matrix Plates for ES Cell Culture, you get:

• Only one cell type to maintain: your ES cells

• Ready-to-use convenience

• An optimised protocol for standardising your cultures

• Easier DNA, RNA and protein isolation from ES cells

• Improved ES cell transfection efficiencies

• Compatibility with multiple ES cell lines and media conditions

• A Quality Assurance tested product for guaranteed lot-to-lotconsistency

A feeder-free system

In 2001, the first feeder-free hES culturesystem was demonstrated.1 It has indeed beenshown that hES cells can be grown in theabsence of feeder cells, as long as they areplated on an appropriate extracellular matrix(ECM) and cultured in feeder-conditionedmedia (MEF-CM) or media supplemented withappropriate soluble factors. BD MatrigelTM

Matrix, coupled with different conditioned ordefined media, is now widely accepted as analternative substrate for culturing hES cells.2-4

a) b) c)



As a leading provider of integrated cell andtissue culture systems for selected cell typesand applications, BD Biosciences – DiscoveryLabware continues to break barriers bybringing you a comprehensive selection oftools and solutions for feeder-free hES cellculture.

With the NEW BD BioCoatTM MatrigelTM

Matrix 6-well Plates for Embryonic Stem(ES) Cell Culture, take the step towardsfeeder-free, streamlined and reproduciblelong-term hES cell cultivation.

BD BioCoatTM MatrigelTM Matrix Plates for ES Cell Culture are 6-well BD FalconTM plates,coated with BD MatrigelTM MatrixTM followingan optimised coating process. Used incombination with appropriate conditioned ordefined media, the BD BioCoatTM MatrigelTM

Matrix Plates for ES Cell Culture allow forstreamlined and reproducible cultivation andexpansion of your ES cells, in a feeder-freesystem.

An optimised protocol

BD BioCoatTM MatrigelTM Matrix Plates for ES Cell Culture were developed using MEF-CM, due to a lack of defined commercialmedia that are qualified for feeder-free ES cultivation. BD Biosciences has optimised aculturing protocol which includes pre-qualified

Cell culture

Product Description Packaging Cat. No.

BDTM bFGF, human recombinant 10 µg 734-1405 50 µg (5 x 10 µg) 734-1406100 µg (10 x 10 µg) 734-1407

BDTM Dispase (5000 caseinolytic units) 100 ml 734-1312

BD FalconTM 12,5 cm2 Canted Neck Flasks with plug-seal cap 100 / case 734-0010

BDTM PuramatrixTM Peptide Hydrogel 5 ml 734-1398

BDTM 3D Collagen Composite Scaffold 24 scaffolds in one 734-1061BD FalconTM 48-well plate

BDTM 3D OPLA® Scaffold 24 scaffolds in one 734-1062BD FalconTM 48-well plate

BDTM 3D Calcium Phosphate Scaffold 24 scaffolds in one 734-1063BD FalconTM 48-well plate

Product Description Packaging Cat. No.

BD BioCoatTM MatrigelTM Matrix 6-well Plates for ES Cell Culture 5 plates 734-1404

Related Products

Figure 2. Immunofluorescent images of H9 cells grown on BD BioCoatTM MatrigelTM Matrix Plates for ES Cell Culture. H9 cells were maintained beyond 7 passages (~ 40 population doublings) andstained positive for expression of well-characterised, undifferentiated markers a) Oct-3/4 and b) SSEA-4 (green; Hoechst nuclear stain: blue). Immunofluorescent images were acquired on a BD Pathway™ Bioimager using a montage feature to capture a larger portion of colonies.

reagents such as BDTM human basic FibroblastGrowth Factor (bFGF) as a media supplement.Using this protocol, H9, H14 and H1 cell lines(from the WiCell Institute) have been culturedon BD BioCoatTM MatrigelTM Matrix Plates for ES Cell Culture with MEF-CM supplementedwith 8 ng/ml bFGF for over 40 populationdoublings.

Performance demonstrated

hES cells cultured on BD BioCoatTM

MatrigelTM Matrix Plates:

• Maintain normal karyotype

• Demonstrate a stable proliferation rate and high telomerase activity

• Maintain the expression of cell markerscharacteristic for undifferentiated hES cells

• Retain pluripotency, and form embryoidbodies (EBs) in vitro and teratomas insevere combined immunodeficient(SCID) mice

• Differentiate into cells from all threegerm layers

BD BioCoatTM MatrigelTM Matrix Plates forES cell culture are suitable for maintainingand expanding multiple ES cell linesincluding H1, H7, H9, H14, HES-NCL1, SA 002,AS 038, SA121 and SA167.Please contact [email protected] forfurther details and references on media andgrowth factor combinations used.

References1. Xu, C., et al., Nature Biotechnolgy 19(10):971 (2001).2. Xu, C., et al., Stem Cells 22(6):972 (2004).3. Stojkovic, P., et al., Stem Cells 23(3): 306 (2005).4. Sjögren-Jansson, E., et al., Developmental Dynamics

233(4):1304 (2005).

a)

b)

BRAND now offers deep-well plates (as well as PCR®- and microplates) with bar codes printeddirectly on the plates. Direct printing makes the bar coding more flexible, rugged and cost-effective than ever before. During the introductory period, BRAND is offering custom printedbar coded plates without any additional charge.

Recently, BRAND converted its offering from printed bar code labels to directly printed plates.BRAND can rely on decades of experience printing laboratory glassware and plastic ware.The high-quality, direct-print process results in a bar code that is significantly more rugged,cost-effective and flexible than applied labels.

A specification sheet that is available at www.brand.de may be used to define your individualbar code printing requirements and to order samples free of charge. Here you can specify, forexample, the type of bar code, the bar code position on the plate, or the plain text position. Basedon your specifications, BRAND experts work closely with you to provide an optimal, individualsolution.

During the introductory period of the BRAND direct printing bar coding system, BRAND is offeringbar coded plates with an individual designation at no additional charge. Plates may also beprinted, for example, with your organisation logo, or even with the abbreviation of a particulardepartment or working group.


Get individual bar codes on your deep-well plates

Capacity (ml) Material well Shape Pack Cat. No.

0,5 PP round 48 391-01581,1 PP round 24 391-57041,1 PS round 32 391-01592,2 PP square 24 391-5705

Deep-well plates, 96-well, U-bottom wells, non-sterile

• 12 x 8 configuration• Capacities 0,5 ml, 1,1 ml and 2,2 ml• Designed for a wide range of applications, such as screening, cell and tissue cultures, serial

dilutions, reagent transfer and sample storage down to -80 °C (PP) or -20 °C (PS) • Deep-well plates are stackable for easy storage

More features of the BRAND deep-well plates include:

• Standard SBS footprint

• Polypropylene for high chemical resistance, e.g., DMSO, phenol, chloroform

• Autoclavable at +121 °C, 20 min, polypropylene (PP)

• Optimal sample mixing and recovery using U-bottom wells

• Alphanumeric code and cut-away corner simplify sample identification and orientation

• Cover mats specially designed to reduce sample evaporation

• Sample storage down to -80 °C, PP, or -20 °C, polystyrene (PS)

Samples of individual bar codes

BRAND moves bar coding from printed labels todirect printing - The difference is in the details!



• PP. 24 x 16 configuration• Capacity 0,3 ml• Designed for a wide range of applications,

such as High-Throughput Screening (HTS),cell and tissue cultures, serial dilutions,reagent transfer and sample storage downto -80 °C

• Centrifugation to 4000 RCF• Deep-well plates are stackable for easy

storage. Pack of 48

Cat. No. 391-5712

Labeling

Description Material Pack Cat. No.

for 0,3 ml 384-well plates Silicone 50 391-0093for 0,5 ml 96-well plates PP 50 391-5713for 1,1 ml 96-well plates mod. PE 24 391-5706for 2,2 ml 96-well plates EVA 24 391-5707

See our promotion in Shop!

• Cover mats reduce the maximum volume ofwells

• Adhesive sealing films can also be used

Deep-well plate, 384-well, U-bottom wells, non-sterile

Cover mats for deep-well plates

OptiCell consists of two parallel gas-permeable, cellculture treated polystyrene membranes 75 µm thick and 2 mm apart. Each side has a growth area of 50 cm2, total100 cm2. The thin profile and microwell footprint designmaximises incubator space and reduces the consumptionof medium. The recommended volumes are 10 ml forOptiCell and 30 ml for OptiCell MAX. Access to the cells isvia two resealing ports providing a closed growthenvironment with sterile fluid path, minimising the risk ofcontamination. OptiCell is barcoded for easy tracking andautomated handling.

A full range of accessories are available:The OptiCell Rack holds 20 OptiCell or 10 OptiCell MAX Chambers. Multiple racks can be connectedtogether. Filling and other manipulations are facilitated by the single use OptiCell Tip. This has astainless steel 18 gauge blunt tip and is both sterile and non-pyrogenic, fitting standard syringes.The OptiCell Knife can be used to remove growth membranes from the frame for sectioning. Formagnetic separation within the growth chamber, use the OptiMag, a reusable, flexible magnetwhich attachs to OptiCell. For transporting cell cultures use the OptiCell Mailer. The breathable,absorbent sleeves with cardboard mailers ensures the arrival of cells in good condition.

LiveCell Array Microscope slide

The LiveCell Array Microscope slideis an easy-to-use, convenient andinexpensive technology for imagingindividual cells (non-adherent oradherent) under various, controlledconditions. It is compatible withstandard microscopes, and “off-the-shelf” imaging software. The regularspacing of the picolitre sized wells,combined with a 0’0” locationmark, enable researchers to re-locate a specific cell of interest,even if the slide has been removedfrom the microscope for short-termincubation or processing. Cellbiologists can utilise this unique“honey comb” design to examineheterogeneity within cellpopulations and obtain kinetic datafrom asynchronous cells.


Cells are loaded into the OpticalLiveCell Array through the cellapplication well using a standardpipette.

Individual cells settle into the individual wells by sedimentation.

Solutions are applied to the raisedarea, and are drawn via micro channels over the cells by capillaryaction. Displaced solutions accumulate in the waste reservoir.

Nunc OptiCell® cell culture system

Applications*Transportation of live cells OptiCell mailer kit• Safe and convenient - incubator to mailer to

incubator• Access to O2 and stable pH during shipping• Only device approved by IATA as a primary

container

Hybridoma antibody productionOptiCell MAX kit• Completely sealed sterile system• Allows harvesting of antibody 3 times a

week for higher antibody concentration andyield, without centrifugation

Freezing & thawing cells• Effective and convenient short-term

storage of cells at –80 °C and long-termstorage at –152 °C

• As no trypsination is needed, there isquick recovery and growth afterthawing

Biomagnetic cell separationOptiMag kit• Minimise cell manipulation during

magnetic cell separation –separate and continue cellculture in the same device,without centrifugation

• Compatible for use with Dynabead®

antibodies and MACS® antibodies

Cell imaging & staining• Cultures can be fixed and stained in situ,

low background fluorescence• Membrane can be sectioned for small scale

staining

Transfection• No need to change the transfection

reagents or protocols• Simplify the process of isolating single cells

Dynabead and MACS are trademarks of Dynaland Miltenyi Biotech respectively.*Application Notes available

Cell culture samples consisting of multiple cellsprovide only an average response rate tomanipulation. Whole cell populations therebymiss unique responses of cell subpopulations.Scientists could benefit from the ability to studyeach cell individually and observe real-timeresponses to a drug or an experimentalcondition. Analytical technology used in cell-based assays has progressed to the point wherecell populations can be observed at single cellresolution. Advanced high-content analysis(HCA) techniques enable individual andsequential cell measurements, but they requireexpensive instrumentation and can be used onlywith adherent cells. To overcome theselimitations, the first slide based HCA tool tofacilitate the study of individual, living, adhering,and non adhering cells within a cell populationwas developed. In addition, real-time datacollection may be conducted on individual cellsthroughout the process of interventions, such asexposure to drugs or altered experimentalconditions. The NUNCTM LiveCell ArrayTM

microscope slide contains a compact array ofPico wells that are available in four sizes.The 15 micron and 20 micron diameter wellsaccommodate individual cells, such as cell lines,primary blood cells, primary bone marrow cells,and beta cells. The 100 micron and 250 microndiameter wells accommodate large cells orgroups of smaller cells. Common cells for theselarger Pico wells include plant cells, neuronalcells, cell clusters, including islets of langerhansand neurospheres, and tissue-like cellaggregates. The array and its sliding cover slip(Figure 1) are made of glass and embedded in aplastic slide frame. Cells are applied to the arrayand covered with the sliding cover slip. Washsolutions, dyes, and treatments are applied to araised area at the edge of the cover slip, andfluid is drawn by capillary action over the cells.The LiveCell ArrayTM can be used with anystandard upright or inverted microscope andallows scientists to easily perform HCA using aminimal number of cells and costly reagents. Inaddition, it offers the researchers the ability to:• Perform long-term, non-intrusive, and

repeated measurements on intact, living,adherent, or non- adherent cells

• Study heterogeneous cell populations toobserve individual responses to treatmentand rare events

• Perform multiple functional assays on livingcells followed by post fixation studies on thesame cells, as well as correlate between datasets (e.g. intracellular immunoassay)

• Perform kinetic measurements of non-synchronous activities in individual cellsobserving each cell

• Perform measurements on a cellular-to-molecular level, correlating whole cellmeasurements with molecular events

• Analyse and compare actual measurementsof subpopulations and individual cells, ratherthan use mean values of entire populations

Reactive Oxygen Species To illustrate the unique function of the LiveCellArrayTM , selected data from recent findingsdemonstrating temporal measurement of thegeneration of reactive oxygen species (ROS) inidentifiable individual cells was presented. ROSare involved in multiple physiological responsesthrough modulation of signalling pathways.These free radicals and their metabolites induceboth positive and negative cellular effects

through complex metabolic pathways. TheLiveCell ArrayTM was used to simultaneously anddirectly determine their production dynamics andconcentrations in each living cell. PromonocyticU937 cells were stained with 10 µMdihydrorhodamine123 (DHR123), a dye used toassess intracellular levels of ROS, and loadedinto the LiveCell ArrayTM . Each cell occupied asingle picowell and did not change its originallocation upon staining or washing. A 50 µMhydrogen peroxide (H2O2) solution was appliedto the cells to stimulate ROS generation, anddata was obtained before and after a 30 minuteexposure. Figure 2 shows bright field andfluorescence images of U937 promonocytesstained with DHR123 and an SEM micrograph ofU937 cells within the array. Positive staining isobserved in most cells, although intensity is notuniform. The sub cellular distribution of DHR123shows the staining pattern of the mitochondria,which was found to be highly heterogeneouswithin a cell population.



The NUNCTM LiveCell ArrayTM

Cell culture

Multiparametric assays on individual cellsTemporal measurement of reactive oxygen species generation

Sarah Haigh, Ph.D., Dan Schroen, Ph.D., Naomi Zurgil, Ph.D., and Mordechai Deutsch, Ph.D.

Application example

Figure 2. (A) Light and (B) fluorescence micrographs of individual U937 cells stained with 10-µMDHR123 and then incubated for 30 minutes in the presence of 50-µM H2O2. Scalebar=20 µm(C) A scanning electron micrograph of U937 cells within the NUNCTM LiveCell ArrayTM

Figure 1. NUNCTM LiveCellArrayTM microscope slide.Inset: SEM images ofpicowell array, empty and loaded with cells

Pall Life Sciences manufactures membranes and chromatography resins that exhibit high resolutionand binding capacities with low non-specific binding for the purification and concentration ofproteins. We incorporate these media into devices that use fast yet gentle methods and minimisehandling to protect samples.

Pall also offers a variety of products for protein detection including PVDF and nitrocellulosemembranes for western blotting; available as cut membranes or integrated into multi-well filterplates.

Products

➡➡ Chromatography Media for affinity, ion exchange, size exclusion, hydrophobicinteraction and hydroxyapatite chromatography

➡➡ AcroPrepTM and AcroWellTM Filter Plates for superior performance in highthroughput sample preparation and detection procedures

➡➡ Centrifugal Devices facilitate pure, concentrated product with high recoveries

➡➡ Nanosep® MF (microfiltration) Devices for applications such as particulateremoval prior to sample analysis and as a housing to easily manipulate theprocessing of bead-bound targets

➡➡ EnchantTM Protein Purification and Depletion Kits for the depletion,fractionation, and purification of abundant proteins

➡➡ MinimateTM TFF System, a rapid and efficient system for the separation andpurification of biomolecules up to 1 litre

➡➡ Transfer Membranes offer high binding capacity and low background forincreased sensitivity in protein detection

BioSepra resins facilitate high capacity purification

The BioSepra line of chromatography resins greatly simplifies protein purification and fractionation.

Biosepra Ceramic HyperD® ion exchange resins offer high dynamic binding capacities and fastflow rates. By tailoring attributes such as chemistry, pore size and resin diameter to specificapplications, Pall chromatography resins exhibit the highest performance characteristics possiblewhile ensuring reliable, reproducible protein isolation.

Efficient removal of detergents from protein solutionsusing the BioSepra SDR HyperD resinChromatography resins for detergent removal. SDR HyperD resin binds detergents used in viralinactivation processes (e.g. CHAPS, SDS, ASB14). High recovery of proteins (exclusion limit 10 kDa)is obtained. The resin exhibits high adsorption capacity for small hydrophobic molecules and isstable in acidic, polar organic and oxidising solutions.


Pall Life Sciences – accelerate protein samplepreparation and analysis

StartingSample

Deplete

Fractionate/Purify

Concentrate

Desalt/RemoveDetergent

Analyze

Acrodisc® Syringe FiltersVacuum Filters

EnchantTM ProteinPurification Kits

BioSepra® Resins

Centrifugal Devices

AcroPrepTM Filter Plates

Centrifugal Devices

AcroPrep Filter Plates

BioSepra Resins

Blotting Membranes

Protocols for SDR HyperD solvent-detergent removal are available in gravity flow column or spincolumn configurations.

Achieve > 98% removal of albumin and IgG from 50 mlof serum/plasmaThe new Enchant™ Multi-Protein Affinity Separation Kit for the depletion of human serumalbumin (HSA) and/or human IgG. The kit utilises a protein-based specific capture method toachieve maximal depletion of the target proteins and minimal loss of other proteins. Depletion ofHAS or IgG is typically 98%. The ligands have high specificity for the target proteins and can beused under harsh conditions intended to disrupt protein interaction. Depletions take approximately20 minutes from start to finish.


For more information on these products contact your local VWR sales office,send an e-mail to [email protected] or visit our website www.vwr.comSample preparation

Description Pack Cat. No.

SDR HyperD Solvent-Detergent Removal Chromatography ResinSDR HyperD 5 ml 555-0011SDR HyperD 25 ml 555-0010SDR HyperD 100 ml 555-0009Enchant™ Multi-Protein Affinity Separation Kit Pack for 24 Samples 733-1500

Detergent Protein solution Protein solution Protein solutionTriton (DBC = 60-80 mg/ml) IgG AT-III Bovine serum

Initial conc. (ppm) 10.000 10.000 10.000

Final conc. (ppm) < 10 < 10 340

Removal efficiency > 99,9% > 99,9% > 95,2%

For a complete listing of Pall’s Protein Purification product range or toorder a new Protein Sample Preparation and Analysis Application Manualplease contact VWR customer services.

Custom kits for relative and absolute quantification of proteins by LC-MSOne of the key challenges in Proteomics is the quantification of proteins at very low concentrationsin complex protein mixtures.

Result of post-translational modifications is a significantly increased number of proteins of aproteome than the number of genes that constitute the corresponding genome. A need exists fortechnologies that permit the direct quantification of proteins and post-translational modifiedprotein expression levels.

As one of the leading manufacturers of mass spectrometers we have a large base of customersfacing these challenges on a daily basis.

We were also the first company to install a pilot research centre dedicated to Biomarker discovery,the BRIMS centre in Boston, MA.

HeavyPeptidesTM AQUA for absolute quantificationThe isotopic dilution assay development time is significantly shorter than other methods, and it isthe only technique offering absolute protein quantification (AQUA) by MS. Best results can begenerated on a triple quadrupole mass spectrometer such as the Thermo Scientific FinniganTM

QuantumTM family.

Increased flexibilityCarefully listening to end-users led us to be even more flexible to meet your needs.

New Thermo ScientificHeavyPeptideTM

• Up to two phosphorylations per peptide

• Choose between 5 or 40 aliquots per peptide

• Available with more than oneheavy amino acid

• Peptide length up to 30 AA

• LightPeptidesTM (control)


Ultimate flexibility for your protein quantification needs

HeavyPeptidesTM AQUAcustom kits

Peptide• Isotopic labelled peptide or non-labelled

peptide for control purposes (lightPeptide)

Specifications• Purity > 97%• Length: 8-15 aa• Custom kits: 5 vials x 2 nmol (5 pmol/µl)

Optional services• Up to two phosphorylations• Additional heavy amino acids• Custom XL kits: 40 vials x 1 nmol

(5 pmol/µl)• additional AA up to a total length of

30 AA per peptide

Shipment• In solution on wet ice

Quality control• Mass Spectrometry• Analytical HPLC• Quantification by Amino Acid Analysis

(AAA)

FREEdemo kit Please contact your local VWR specialist or email [email protected]

to receive a voucher for your free HeavyPeptide AQUA demo kit.



Flexible and powerfulAs HeavyPeptides can be prepared with covalent modifications (e.g., phosphorylation, methylation,acetylation, etc.) that are chemically identical to naturally occurring posttranslational modifications,they represent one of the most powerful tools for relative and absolute quantification by LC-MS ofproteins prone to these modifications.

Economically viableWith HeavyPeptides available from € 154,50 and a typical experiment requiring less than 100fmole, the cost per experiment is extremely low3.www.thermo.com/heavypeptide

IP and TrademarkAQUA: This method was developed by Dr.Steve Gygi and colleagues at Harvard Medical School[Stemmann O, Zou H, Gerber SA, Gygi SP, Kirschner MW; Dual inhibition of sister chromatid separationat metaphase, Cell 2001, Dec 14, 107:715-726] and is the subject of both US and PCT PatentApplications. Limited non-commercial use of this method is permitted under a licensing arrangementwith Harvard Medical School.AQUA is a trademark of Harvard Medical School. HeavyPeptide, Quantum, Finnigan and BioWorks aretrademarks of Thermo Electron Corp.

Protein quantification

phos.

( )p

Native Protein Phosphorylated Protein

A proteotypic peptide is identified and characterized3

( )p

An ternal standard is synthesized. A stable isotopically

labeled amino acid is incorporated during the process. The

resulting product is named HeavyPeptide or HeavyPeptide

AQUA

in

4

The protein sample is spiked with a known amount of the

HeavyPeptide and digested

The samples are analyzed by LC-MS/MS

phos.

phos.

Protein abundance is derived from MS peak areas5

See our promotion in Shop!

Fields of application are:

Relative quantification- Cell signaling profiling- Expression profiling- Structural studies

Absolute quantification- Biomarker validation- Diagnostic - Allergenes quantification in food processing

AQUA mass spectrometry quantification meets the following criteria:

• Accuracy – absolute quantification of proteins in complex samples

• Specificity – custom-made product ensures ultimate specificity foryour quantification assay

• Sensitivity – low abundance protein quantification

This paper shows how the Thermo Scientific Multiskan®

Spectrum microplatespectrophotometer together withThermo Scientific Microtiter® UVMicroplates can be used for DNAand protein analysis with excellentsensitivity. The detection limit for DNAquantitation in Thermo Scientific384-well UV Microplatescalculated according to the IUPACstandard 3SD method is 60 ng/ml.For BSA quantitation in ThermoScientific 96-well UV Microplatesthe detection limit is 7 µg/ml.After pathlength correction thedetection limit is 4 µg/ml. Spectralanalysis of DNA in 96-well UVMicroplates is possible withsamples clearly smaller than 2 µgof DNA.The stable and low background ofboth instrument and microplatesenables the measurement of smallsample concentrations and hencesaves sample volumes.


High sensitivity DNA and protein analysis with Thermo ScientificMultiskan® Spectrum and Microtiter® UV microplates

Nucleic acid and protein quantitation with highsensitivity is needed in a wide variety ofbiological applications. Nucleic acidconcentration is commonly measured bydetermining UV absorbance at 260 nm. Forconcentration determination of protein solution,UV absorbance at 280 nm is measured.The absorption of nucleic acid bases adenineand guanine have an absorbance maximumslightly below 260 nm and thymine andcytosine slightly above 260 nm. The absorbancepeak of nucleic acids depends on the basecomposition.The absorption of proteins at 280 nm is mainlydue to the aromatic amino acids tyrosine andtryptophan and disulfide bonds of cystine (1).The relationship of absorbance to nucleic acidor protein concentration is linear, following theBeer-Lambert law: A = εxCxl, where A = theabsorbance, e = the solution’s extinctioncoefficient, C = concentration and thepathlength of light.Absorbance measurements are fast andconvenient, since no additional reagents orincubations are required. Also, the sample is notconsumed and may be used for further analysis.Traditionally these measurements have beenperformed in quartz cuvettes, but nowadays it ispossible to use quartz microplates or disposableUV transparent microplates. The advantage ofmicroplates is that more than one sample atonce can be measured which enables higherthroughput. With disposable UV microplatesthere is no need for washing and there is norisk of sample carry-over as with reusablequartz microplates or cuvettes.

In this application note we present examples ofDNA and protein measurements with theMultiskan® Spectrum UV/Vis spectrophotometerand Microtiter® UV Microplates.

Materials and methods

DNA from calf thymus was diluted in distilledwater to concentrations between 1,25 and 100 µg/ml. Protein solutions withconcentrations between 25 and 1000 µg/mlwere prepared from bovine serum albumin(BSA) in distilled water.200 µl of each dilution in four replicates wasadded to wells of 96-well UV Microtiter®

Microplates and 50 µl in four replicates to 384-well UV Microtiter® Microplates.Absorbance measurements were performedwith Thermo Scientific Multiskan® Spectrum (Cat. No. 735-0263).

Results and discussion

A DNA standard curve for concentrationsbetween 1,25 and 100 µg/ml is shown in Figure 1. Measurements were performed withsample volumes of 50 µl in 384-well UVtransparent Microtiter® Microplates. Thestandard curve is linear at a broad range ofconcentrations enabling DNA quantitation ofsmall sample volumes at low and highconcentrations. The detection limit for DNAquantitation calculated according to the IUPACstandard 3SD method is 60 ng/ml indicating thesensitivity of this measurement.



Figure 2 shows BSA standard curves forconcentrations between 25 and 1000 µg/ml.Measurements were performed with samplevolumes of 200 µl in 96-well UV transparentMicrotiter® Microplates. Also here linearity overa broad concentration range is observed.With Multiskan® Spectrum SkanIt® Software itis possible to correct for the pathlength oflight, making the results obtained withmicroplates directly comparable with data fromcuvette measurements.The detection limit for BSA quantitationcalculated according to the IUPAC standard3SD method is 7 µg/ml. After pathlengthcorrection the detection limit is 4 µg/ml. Thepathlength correction function takes the liquidlevel into account and so minimises the effectof pipetting errors. This explains the smalldifference in detection limit.Absorbance spectra of DNA solutions withconcentrations between 1,25 µg/ml and 10µg/ml were measured in 96-well UVtransparent Microtiter® Microplates (Figure 3).The figure shows that spectra can be measuredfrom samples clearly smaller than 2 µg of DNA.

References1. Pace, C.N. et al. 1995. How to measure andpredict the molar absorption coefficient of aprotein. Protein Sci 4, 2411-2423.

DNA and protein analysis

Figure 1. DNA standard curve in 384-well UVtransparent Microplates.

Figure 2. BSA standard curves in 96-well UVtransparent Microplates. The graph shows theresults as such and after applying pathlengthcorrection.

Figure 3. Absorbance spectra of DNA at concentrations between 1,25 and 10 µg/ml.Measurements were performed in 96-well UVtransparent Microplates. Sample volumes were200 µl/well.

Multiskan® Spectrum

Description Cat. No.

Multiskan® Spectrum with cuvette, SkanIt Software Drug Discovery Edition 735-0287

Multiskan® Spectrum with cuvette, SkanIt Software Research Edition 735-0263

Multiskan® Spectrum without cuvette, SkanIt Software Drug Discovery Edition 735-0264

Multiskan® Spectrum without cuvette, SkanIt Software Research Edition 735-0265

Fig

ure

1.

Fig

ure

2.

Fig

ure

3.

The KrosFlo® Research II TFF System is ideal for performing and monitoring the filtration of volumesranging from 2 ml to 500 ml. Whether maintaining a validated separation or characterising aprocess for scale-up, this system combines process surveillance with flexibility for a multitude ofapplications. Operated by a 2,3 lmin-1 peristaltic pump, the KrosFlo hollow fiber module quickly andefficiently filters while the disposable flow-path minimises hold-up volume. The system features theKrosFlo Digital Pressure Monitor that reads and displays process pressures with alarms that canshut-off the pump under high or low-pressure conditions. Great for validated processes andprotecting valuable samples! The monitor also interfaces with a PC to record all process data intothe included KF Comm data collection software. Depending on your process volume or the moduletype, select from 2 system models available, let the KrosFlo Research II TFF System efficientlyprocess and protect your research sample

KF Comm data collection softwareInterface with a PC to download real-time operating pressures, calculated TMP, pump rate, date andtriggered alarm points into this specially designed Excel® format software for easy collection andgraphing of process data. Measured flux rates can be manually entered along with the streamingdata feed.

KrosFlo® digital pressure monitorKeeps a "watchful" eye on your process separation by using 3 pressure transducers. In addition todisplaying 4 readings, the monitor can be set with 5 alarm points, 2 of which stop the pump toprotect the sample.

Inlet pressure Permeate PressureHigh stop alarm Low warning alarmHigh warning alarm Low stop alarmLow stop alarm


The NEW KrosFlo® Research II TFF system

Tangential flow filtrationsystem for volumes of 2 ml to 500 ml


For more information on these products contact your local VWR sales office,send an e-mail to [email protected] or visit our website www.vwr.comFiltration

Description Cat. No.

KrosFlo® Research II TFF SystemsKrosFlo®, flow-path type: MicroKro® with C-flex 14, 130 ml/min 515-0207KrosFlo®, flow-path type: MidiKros® with C-flex 16, 480 ml/min 515-0206

System ComponentsKrosFlo® pressure monitor, 220 V; KF Comm, 3 transducers & RS232 515-0200Disposable pressure transducers for KrosFlo® pressure monitor (3/pkg) 515-0201KrosFlo® Research II pump, 2.3 ltr/min, 220V (pump head not included) 515-0202KrosFlo® Research II pump head, 3 SS rollers 515-0204MicroKros® Flow-path fitting kit (size 14 tubing & fitting) 515-0205MidiKros® Flow-path fitting kit (size 16 tubing & fittings) 515-0203

Research scale applications

• Cell washing

• Lysate clarification

• Bacterial concentration

• Micro-particle diafiltration

• Macromolecular isolation

• Virus purification

• Viral clarification

• Protein purification

• Antibody production

• Desalting & buffer exchange

Please enquire and we'll help you find theKrosFlo® Research II TFFsystem that best suitsyour application

KrosFlo® Research II pumpProvides precision flow control along with predictive flow rates for standard MasterFlex® tubingand calibration for non-standard tubing. The 2,3 lmin-1 peristaltic pump can also be programmed forpriming and dispensing. The pump head adjusts to 4 different orientations. For maximum utility, twopump heads can be operated simultaneously.

System design: stability & flexibilityThe stand that supports the process flow path is integrated with the pump to provide ultimatesystem stability. The module and reservoirs are secured in place by "ready-lok" holders that easilyattach, adjust and swivel on a pair of rails positioned on both sides of the pump, providingcomplete flow-path flexibility. The pressure monitor can either remain separate or be secured to thetop of the pump.

Module specificationsMembrane type Fiber ID MWCOMixed Cellulose Ester (ME): 0,6 mm 0,1 µm – 0,2 µm

Polyethersulfone (PES): 0,5 mm 0,2 µm – 0,5 µmPolysulfone (PS): 0,5 mm 0,05 µm – 400 kD – 100 kD – 50 kD – 10 kD

1,0 mm 500 kD – 100 kD>


MIEF-SYS Isoelectric Focusing horizontal electrophoresis unit

New! The new Isoelectric Focusing (IEF) unit has been designed and optimised for applications usingeither ready-to-use pre-cast or hand-cast polyacrylamide gels, as well as for Immobilised pHGradient strips (IPG) strips used in high resolution 2D gel electrophoresis of proteins.

Numerous flat bed electrophoresis applications can be performed up to a maximum 1500 Volts inone of three modes:

• Wick-based electrophoresis

• Pre-cast or hand-cast horizontal gels

• IEF on dried gel strips with an immobilised pH gradient (IPG strips)

• Ceramic Cooling Plate - 27 x 27 cm (W x L) can support polyacrylamide slab gels: 2 x (10 x 10 cm) or 1 x (20 x 20 cm) or up to 30 IPG Strips 3 mm wide x 24 cm long

• Ceramic Cooling Plate can be connected to any circulating water bath to maintain IEF at 4 °C for polyacrylamide gels or 20 °C for IPG strips

• Acrylic Electrode Frame - sits directly on the ceramic coolingplate for use with IEF polyacrylamide gels

• Glass Electrode Frame - sits flush with the ceramic coolingplate for optimal cooling efficiency and minimises themess caused by oil application required during IEF with IPGstrips

• Positive and Negative Electrodes made from clear acrylic clipneatly within the acrylic or glass electrode frame, allowing the voltage gradient to be fine-tuned along the entire length of each IPG strip

• Glass Electrode Weight - ensures that the electrodes remain in complete contact with the positive and negative ends of each IPG strip or IEF gel during IEF

• 2 mm shrouded power output connectors, compatible with most high voltage power supplies

• The IEF tank and cooling plate can be used for wick-based horizontal gel electrophoresis with any polyacrylamide gel

BEN

EFIT

S

Outline protocol for IEF gels

1. Place the cooling plate in position within the tank. Set the cooling temperature of the circulator chiller,connected to the cooling plate by quick-fit tubing, to 4 ºC at least 10 minutes before using the unit.

2. Add 200 µl of Bayol F or kerosene onto the cooling plate, and spread evenly over an area roughly the samesize as the IEF gel, to optimise heat exchange.

3. Place the gel directly on top of the area covered by Bayol F or kerosene and, by using a pipette orelectrophoresis roller, gently disperse any bubbles underneath the gel. Remove with a tissue any surplus BayolF or kerosene, as both chemicals are flammable.

4. Immerse the 17 mm filter paper electrode wicks in their respective electrode solutions, depending on whetherit is the anode or the cathode - i.e. the anode electrode wick will require a lower pH solution, while a higherpH solution is necessary for the cathode electrode wick. The length of the electrode wicks should correspondto the width of the IEF gel, otherwise short circuits or band distortion may result.

5. Remove the any protective film covering the IEF gel and position the electrode wicks at either end of the gel,at least 5 mm inside each edge. The distance between each electrode should be 10 cm for most commerciallyavailable IEF gels.

6. Place a sample applicator strip in the middle of the gel, after washing it free of any dust or debris withdistilled water.

7. Load 10 µl of protein sample into each well with a pipette, reserving at least one well for a 10 µl-aliquot ofIEF Markers 3-10.

8. After loading the gel, carefully lower the glass electrode weight over the anode and cathode electrodes, sothat they remain in contact with the electrode wicks and gel during the application.

9. Perform IEF according to the recommended running conditions in the user manual. Once IEF is complete, theIEF gel is ready for staining.

Outline protocol for IPG strips

1. Incubate IPG strips overnight with buffer (8M Urea, 1% CHAPS, 13 mM DTT and 0,5% Ampholyte 3-10,corresponding to the pH gradient of the IPG strip). At least 5 and 50 µg of protein is sufficient for 7 and 18 cmIPG strips respectively.

2. Overlay each strip with silicone oil to prevent desiccation during the overnight incubation period.

3. Attach the chiller unit, preset at 20 °C, to the ceramic cooling plate of the MIEF-SYS unit using the quick-fitconnectors.

4. Place the glass electrode frame directly on the cooling plate and apply a thin layer of silicone oil over theglass plate before laying out the IPG strips, allowing a minimal distance of 3 mm between each strip.

5. Cover the anode and cathode ends of the IPG strips with an electrode wick saturated with deionised water,before clipping the anode and cathode electrodes into position within the glass electrode frame. Overlay theglass electrode weight to keep the electrodes in position.

6. Overlay more silicone oil to prevent urea crystallisation caused by the heat generated during IEF performedaccording to the settings laid out below for 18 cm IPG strips as an example.

7. After IEF incubate the IPG strips for 10 minutes in Equilibration Buffer 1 (50 mM Tris-HCl, pH 8,8, 6 M Urea,30% glycerol, 2% SDS, 0,01% bromophenol blue and 1% (w/v) DTT) by placing the rehydration tray on to anagitating platform.

8. Incubate the IPG strips for a further 10 minutes in Equilibration Buffer 2 (50 mM Tris-HCl, pH 8,8, 6 M Urea,30% glycerol, 2% SDS, 0,01% bromophenol blue and 5% (w/v) iodoacetamide) as described in step 7.

9. Dip each strip briefly in 1 x Laemmli Buffer before overlaying each strip by its 1 mm thick edge along the topof a pre-made 10% acrylamide gel to optimise resolution.

10. Cover each strip with 0,5% agarose solution, allowing it to set before performing second-dimension SDS-PAGE.


For more information on these products contact your local VWR sales office,send an e-mail to [email protected] or visit our website www.vwr.comElectrophoresis

Protein lysate samples run ona wide range pH 3-10 IEF gel

a. Animal Protein Lysates

b. Potato Protein Lysates

Voltage step 1 2 3 4 5 6 end of run

Voltage (V) 150 300 600 1500 3000 330

Time (h) 0,5 0,5 0,5 0,5 2,5 <20

Volt-hours 75 150 300 750 7500 -

New to VWR bioMarke

programme

Highlights of the 5 PRIME product portfolio

The new dimension in PCR®

The self adjusting Mg2+ buffer technology minimises the need for PCR® optimisationAn innovative PCR® buffer technology accelerates the performance of all 5 PRIME PCR® enzymes.A weak chelator provides the optimal Mg2+ concentration during PCR®. Minimal PCR® optimisationis required and saves time and effort.

Innovative Hot-Start/Cold StopTechnology for highly specific Hot-Start PCR®

Improved specificity without enzymeactivation makes Hot-start PCR® fast andreproducible. A non-protein inhibitor blocksreversibly the Taq Polymerase in it’s activesite at lower temperatures and preventsmispriming. At elevated temperature theinhibitor is released from the enzyme, soamplification in each cycle is as specific aspossible.

Ready-to-use RealMasterMixes for probe-based or SYBR Green real-time PCR®

detectionThe combination of the self adjusting Mg2+ buffer with Hot-Start/Cold-Stop technology improvesreal-time PCR® results. Convenient reaction set-up with ready-to-use master mix saves time andminimises errors. This guarantees an excellent real-time PCR® performance every time.

For any nucleic acid from any sample – choosefrom 5 PRIME exciting purification tools

FastPlasmid Mini Kits – no one isolates plasmid faster (9 min)An innovative technology that works with a true one-step lysis for extremely simplified handlingand enormous time saving. Plasmid purification can be performed in as little as 9 minutes. A simpleone-step cell lysis step followed by a bind wash and elute procedure on a column guarantees highquality plasmid that can be used in demanding downstream applications.

PerfectPure RNA Purification System – fast & efficient column-based RNA purificationThe novel column-based technology eliminates the most common difficulties of RNA purification suchas overloading and clogging. This fast and efficient process produces stable, high quality RNA fromwhole blood, cultured cells, and tissues, free of genomic DNA, proteins and other enzymatic inhibitors.

Convenience, ease-of-use and time savings –this is 5 PRIME’s mission to make life easierfor you in your laboratory. To find out more about our exciting solutions contact your localVWR sales organisation or email [email protected]


Your prime partner for perfectresearch

Reagents

Who are we?

5 PRIME is a company founded in2006 by the entrepreneurial spiritof some life science managers. This newly founded companyacquired certain assets and rightsof the Eppendorf 5-Prime reagentportfolio as well as a selection ofproducts from the Gentra productportfolio from QIAGEN.

Products from 5 PRIME areproduced under ISO qualitystandards and the product qualitymaintains the same excellentperformance specifications thatcustomers rely on from Eppendorf5-Prime and Gentra products.

5 PRIME stands for high productquality at a competitive and fairprice.

>As the one source for your everydaylaboratory research the productportfolio spans all common molecularbiology applications such as:

• Nucleic acid purification

• PCR®

• Transfection

• Protein purification

• Molecular biology reagents



New Blotting Sandwiches from Whatman consist of one sheet of transfer membrane and two sheetsof a blotting paper.

The pre-cut membrane with papers in a sandwich format matches the most commonly used pre-cast electrophoresis gels. These blotting sandwiches offer convenience and time savings forhigh throughput laboratories using chemiluminescent detection or other common detectionmethods.

The ‘ready to use’ sandwiches are offered with Protran®, which is made of 100% nitrocellulose, orWestran® PVDF membranes.

Protran® is a UNIVERSAL blotting membrane, which is suitable for Western (protein), Southern(DNA) and Northern (RNA) blots, and has a very high binding capacity due to its microporousstructure. Protran® is hydrophilic, therefore no pre-wetting with toxic methanol is required, and issuitable for all common detection methods (radioactive, colourimetric, fluorescent) as well aschemiluminescent detection based on horseradish peroxidase systems such asECL/SuperSignal/Luminol and all common staining methods.

Westran® CS protein blotting membrane and Westran® S protein sequencing membrane are bothmade of hydrophobic PVDF. They are an excellent choice for protein transfer due to their highaffinity to proteins and their broad compatibility with detection and analysis systems. Westran® CS(Clear Signal) is specifically designed for Western blotting and protein dot-blotting applications.The membrane exhibits high protein capacity and increased signal over a wide range of molecularweights. Westran® S has a very high protein binding capacity, an excellent chemical resistance andis ideal for protein sequencing. It is also compatible for use in non-sequencing Western Blottingapplications.

Blotting membranes

Speed your throughput with time saving WhatmanBlotting Sandwiches

Description Dimensions (cm) Cat. No.

Protran® BA85 / 3MM 7 x 8,5 732-4355Protran® BA85 / 3MM 8,5 x 13,5 732-4356Protran® BA85 / 3MM 7,3 x 8,3 732-4357Protran® BA83 / 3MM 7 x 8,5 732-4358Protran® BA83 / 3MM 8,5 x 13,5 732-4359Westran® S / 3MM 7 x 8,5 732-4360Westran® S / 3MM 8,5 x 13,5 732-4361Westran® CS / 3MM 7 x 8,5 732-4362Westran® CS / 3MM 8,5 x 13,5 732-4363

Available Blotting Sandwiches – supplied in packs of 20

>Key Advantages

• Available in a convenient‘ready to use’ format

• Time saving for highthroughput laboratories

See our promotion in Shop!Buy one pack of blotting sandwichesand get a SUDOKU game free!

Advanced experiments in gene technologydemonstrate that even small amounts of freeDNA molecules are sufficient to causeinfections, recombination or biologicaltransformations [1,2]. The completedecontamination of equipment and surfacesfrom any DNA molecules is important forbiological containment and safety, as well asfor preventing artifacts in PCR® amplificationexperiments. Using new methods to detectextremely low levels of DNA molecules, weinvestigated the molecular mechanism ofaction of various commercially available DNAdecontamination reagents and found that withhigh concentrations of DNA and shortincubation times none of the conventionalreagents destroyed DNA molecules efficientlydespite their corrosive or even toxic properties.AppliChem have developed the new andunique decontamination reagent,DNA-ExitusPlusTM, and in this paper it iscompared with other conventional productsdemonstrating it’s fast and efficient destructionof nucleic acids that is gentle on you, yourequipment and the environment.

Decontamination reagents use differentcombinations of three molecular principles fordestruction or inactivation of genetic material.Modification and denaturation mask but do notdestroy the genetic information encoded inDNA strands and there is a risk that they maybe chemically re-activated. Safe DNAdecontamination depends on the degradationof DNA into very small fragments. Figures 1 and2 show the results of sensitive quantification ofthe fragmentation process by the novel DNA-ExitusPlusTM and conventional agents.No degraded DNA pieces were found whenproducts relied on modification or denaturationmethodology. So based on our currentknowledge of gene technology and theprinciples of recombination, we concluded thatthese reagents are no longer sufficient. Even

reagents that did show degradation of DNAcause only partial destruction and some verylarge DNA fragments that contain the completegenetic information, still survived treatment(Figure 1).

Only Applichem’s DNA-ExitusPlusTM achievedrapid and efficient degradation with it’s strandbreaking activity independent of the size andsequence of the DNA fragments since it worksby chemical action and not an enzymaticactivity. To verify this efficient degradation ofDNA molecules by DNA-ExitusPlusTM, PCR®

analysis was used (Figure 3) proving that noamplifiable DNA templates are present.Defined DNA samples were dried on the innersurface of reaction tubes, followed by treatmentwith DNA-ExitusPlusTM. Many different non-standardised PCR® tests are used todemonstrate successful DNA decontaminationbut with large DNA control templates, low DNAconcentrations, and high dilutions in thewashing steps, evidence is very limited. Only by using PCR® analysis in combination with asensitive DNA degradation test can one be surethat the DNA is only modified or masked andmay cause difficulties later on. It is clear thatspraying DNA-ExitusPlusTM on lab surfacesensures complete decontamination.

Another severe disadvantage of conventionalreagents is revealed in a test for their corrosivepotential. For this purpose different metalplates were incubated with aliquots of theproducts. Figure 4 shows that all conventionalproducts contain aggressive chemicals withcorrosive, harmful or even toxic effectsincluding ingredients such as azides, mineralicacids like phosphoric acid or hydrochloric acid,aggressive peroxides or strong alkalinesubstances like sodium hydroxide. Even afteronly 20 minutes of incubation irreversibledamage of metal surfaces are observed, butDNA-ExitusPlusTM shows no metal corrosion or


New insights into DNA decontaminationDr. Wolfram H. Marx (AppliChem GmbH, Darmstadt)Dr. Karlheinz Esser (multiBIND biotec GmbH, Dortmund)Prof. Dr. Thomas Lisowsky (multiBIND biotec GmbH, Dortmund)

The first reagent really knocking out DNAcontamination: DNA-ExitusPlusTM

Figure 1. DNA degradation by selected conventionalDNA decontamination reagents in comparison withDNA-ExitusPlusTM. Sample preparation:- 200 ng CCC plasmid DNA (7kb) in 10 ml H2O

+ 5 ml sample - Incubation 3 or 10 min. at 20 °C - Denaturation for 3 min. at 92 °C- 200 ng DNA sample per laneC: Control 200 ng CCC plasmid DNA in 10 ml H2O + 5 ml H2O; M: Marker 1Kb Ladder; X1, X2, X3, X4 =competitor's products; D = conventional DNA-Exitus;D+ = DNA-ExitusPlusTM.

Figure 2. Degradation of small DNA fragments byDNA-ExitusPlusTM. To test the degradation of smallfragments, 500 ng of a 750 bp PCR® product and thecorresponding primer were treated for the indicatedtime (1, 2, and 5 minutes) with DNA-ExitusPlusTM. + 5 µlDNA with 5 µl DNA-ExitusPlusTM; C Control 5 µl DNAwith 5 µl water; M molecular weight marker 1 kbladder. After the treatment, the DNA was denatured for 2 minutes at 95 °C.

Figure 3. PCR® test for the complete removal of DNA contaminations by DNA-ExitusPlusTM.Test DNA (0,1 to 1 ng) was lyophilised on the inner surface of PCR® tubes, incubated for 20 secs with sterile water or DNA-ExitusPlusTM, then washed twice with 100 µl of sterile water. For the PCR® test 50 µl of each reaction mixtures were takencontaining primers for the amplification of the control and test DNA. 1 ng of control DNA in each sample proves that the PCR®

reaction is not inhibited so that amplification of a DNA band corresponding to the test DNA indicates that intact DNA molecules are present. If the test DNA is completely degraded the PCR® reaction should not amplify any DNA fragment for thistemplate. The negative control with sterile water (H2O) exhibits DNA bands for the test and control templates whilst after treatment with DNA-ExitusPlusTM only the fragment of the control DNA is amplified.

indeed in other tests (data not shown) nodamage to many different plastic surfaceseither. DNA-ExitusPlusTM therefore offers agentle and environmentally safe alternativethat degrades and removes all DNA moleculeswith high efficiency but is also neither toxic norcorrosive.

Finally, autoclaving is believed to be aneffective method for DNA decontaminationalthough limited to use with heat-resistantmaterials and equipment that fit into theequipment. Under the standard autoclavingconditions, DNA molecules are degraded intofragments of 20 to 30 base pairs and PCR®

analysis demonstrates that even afterautoclaving, larger DNA fragments can beidentified [1], especially when nucleic acids areprotected by protein envelopes (e. g. viruses) orwithin microorganism (e.g. bacteria). Thereaction time of DNA-ExitusPlusTM correspondsto an autoclave run time drying within 10 to 20minutes after spraying on a surface. Due to itschemical composition, DNA-ExitusPlusTM is notheat-sensitive and does not contain volatileand harmful ingredients. Figures 5 and 6 showthe results of comparing it’s effects on bacterialcultures and nucleic acids at elevatedtemperatures to autoclaves. Only the additionof DNA-ExitusPlusTM leads to an efficientdegradation of bacterial DNA, while under thestandard conditions (aqueous solutions,medium) the controls are always positive interms of undegraded / partially degraded DNA.

SummaryIt is clear that performing a PCR® test alone toprove the complete removal of DNA moleculesis not sufficient as such a PCR® test will benegative for decontamination reagents that

modify or mask DNA, whilst failing to removeor destroy it. The true determination of thedecontamination potential of a reagentrequires PCR® analysis in combination with aDNA degradation test. In addition the use ofautoclaving has to be re-evaluated, since thelatest data show that DNA from viruses andmicroorganisms are not inactivated properly.

These are the outstanding and uniquecharacteristics of DNA-ExitusPlusTM:I. Catalytic and cooperative effects of

the components cause a very rapid non-enzymatic, non-sequence-specificdegradation of DNA and RNA molecules

II. All components of DNA-ExitusPlusTM arereadily bio-degradable and not harmful ortoxic for humans

III. No aggressive mineralic acids or alkalinesubstances are used. Equipment andmaterials are not damaged or corroded even after prolonged incubation times

IV. No harmful aerosols when spraying onsurfaces.

Literature:[1] Elhafi, G. et al. (2004) Microwave or autoclave treatments destroy the infectivity of infectious bronchitisvirus and avian pneumovirus but allow detection byreverse transcriptase-polymerase chain reaction. AvianPathology 33, 3003-306.

[2]Burns, P.A. et al.(1991) Transformation of mouse skinendothelial cells in vivo by direct application of plasmidDNA encoding the human T24 H-ras oncogene.Oncogene 6(11), 1973-1978.


For more information on these products contact your local VWR sales office,send an e-mail to [email protected] or visit our website www.vwr.comDNA decontamination

Figure 4. Corrosive potential of selectedconventional DNA decontamination reagents incomparison with DNA-ExitusPlusTM. Metal platesrepresenting typical laboratory materials andequipment where treated with 10 ml of each indicatedreagent for 20 minutes. 0 = sterile water; X2, X3 =competitor's products; D = conventional DNA-Exitus;D+ = DNA-ExitusPlusTM. In some cases one observes apolishing effect by the removal of dirt or oxide layers.

Figure 5. Autoclaving of recombinant bacterialeads to partial DNA degradation. Equal volumesof either water (-) or DNA-ExitusPlusTM (+), respectively,where added to 50 ml cultures of recombinant E. coliand autoclaved at 120 °C and 1,2 bar for 20 minutes.After autoclaving 10 ml aliquots of these cultures whereloaded onto an analytical agarose gel. Please note theDNA degradation by DNA-ExitusPlusTM (+) resulting infragments smaller than 20 base pairs.

Figure 6. PCR® analysis of the autoclaved E. coli cultures from Fig 2. The recombinant E. coli cultures contained a plasmid bearing theampicillin resistance gene (AmpR-Gen). Aliquots (2 ml) of the cultures where analysed by PCR® for the presence of the complete AmpR gene. (-) = E. coliplus water; (+) E. coli plus DNA-ExitusPlusTM;(K) = E. coli plus DNA-ExitusPlusTM plus 2 ng templatefor the AmpR gene (positive control to prove thatPCR® is working); (M) molecular weight marker.

In support of these growing demands, the embedded Web Server, a newnetwork communication tool, has been developed to support the recording andconsultation of data generated from the bio-cryogenic storage tanks.

CryoView is a new generation of electronic device, which has been specially developed to cover the needs in terms of data consultations. Theseare generated by the electronic devices of the cryogenic vessel (temperature,level of liquid nitrogen, events etc…). All these data are placed in CryoView’s own embedded Web Server, every tenminutes for 366 days. It also records 4096 events.

Through its international output connection (RJ45), which is used to connect itto your local Network, all generated data can be consulted with a standardbrowser (Internet Explorer, Netscape, FireFox...). This means that CryoView is a Plug and Play system.

One international single cable, no software to install.


TraceabilityThe level and detail of traceability

in today’s laboratory & health

markets is a top priority.

Everything linked to stored

biological samples, temperature

history, nitrogen levels, alarms,

actions… needs to be tracked.



What can you check?

Cryo preservation

Level and temperature(practically in real time)

Daily report and graphof temperature

Alarms and states(solenoid valve)

Thresholds and alarms definition

Events

Settings:clock, parameters

Activate the openingof the solenoid valve

Available for the RCB, Espaceand Arpege range.(Either in liquid or gaseous phase)


New Tecan compatible tipsCompatible with - Tecan Genesis, Tecan Freedom, Tecan EVO, Tecan Miniprep, Cavro workstations, Perkin-Elmer/Packard Multiprobe II HT and Multiprobe II HT EXwith Hanging Tip workstations.

For more information contact your local VWR Sales Office or [email protected]

In-house mould makingEngineering team has tremendous expertise in designing/ creating moulds that produceconsistent, high quality products.

Automated assemblyNon-sterile product certified nucleic acid and nuclease safe. Product assembledimmediately and placed into stock to reduce delivery times and ensure virtually no back-orders.

Electronic monitoring of manufacturingAdvanced technology supported by experienced personnel results in higher qualityproducts.

Unique technologyAvailable in Maxymum Recovery to further improve accuracy and precision while reducingsample and reagent wastage.

Customer-site testing of new productsTips work on the specified workstations.

Lot-by-lot QC testingThe most complete lot-by-lot testing in our industry ensures higher product consistency.

Extremely broad product offeringAxygen routinely offers tips not offered by the robotic workstation manufacturer.

Tecan compatible tips available in both Liquid Level Sensing and Tecan-style Clear Tips – 20 µl,50 µl, 200 µl, 1000 µl – non sterile, sterile and filter tips.

Axygen products are manufactured in a clean, controlled “state of the art” facility and certifiedRNase-, DNase- and endotoxin-safe on a lot by lot basis.

Reasons to choose Axygen robotic tips


For more information on these products contact your local VWR sales office,send an e-mail to [email protected] or visit our website www.vwr.comRobotic tips

Axygen launch their new pipette tip refill system –

save time, save space, save money, reduce waste, convenient and easy to use.

New products - tip refill system

• Translucent rack allows tips to be easily seen

• Tip insert ‘clicks’ into rack for added product security

• Colour coded insert allows for easy identification of tip

• Both the 10 µl and 200 µl tips can be used directly from the packaging saving time and precious space

• Graduated 200 µl tip to allow for visual checking of volume drawn

• Certified RNase-, DNase- free by lot number - documentation available via Axygen web-site

Special introductory offer see

Shop!

C.B.S. Scientific is proud to introduce the uniquely designed, multipurpose, easy-to-use Dual CoolSystem. This mini-vertical apparatus can accommodate both slab gel electrophoresis and Westernblotting within the same unit without the purchase of additional components. You can run 2 gelssimultaneously under identical temperature controlled buffer conditions for either application. Thismoulded system is designed primarily for use with a variety of pre-cast gels ranging in size from10 x 10 cm to 10 x 8 cm but hand-cast gels can also be run with the use of accessory combs,plates and spacers.

Simple assembly and disassembly:

Assembly: Place gel plates or blotting cassettes in place, close hinged doors and secure latches.Place freezer blocks in lower buffer reservoir receptacles, slide colour-coded core into bottom reservoir, and attach safety cover.

Disassembly: Remove safety cover. Gel or blotting cassettes can be extracted from the core withoutremoving the core from the tank, allowing for quick processing of samples.

It’s unique!

• Run 2 slab gels or 2 electrophoretic transfers in the same unit • Hinged core design for easy loading and removal of gel or blotting cassettes • Freezer blocks fit into compartments in side of unit for optional cooling• Electrophoretic transfers do not require the purchase of special transfer modules• Gel or blotting cassettes can be extracted without removing the core from the tank, allowing for

quick processing of samples and recycling of buffer for the next set of samples• Colour-coded anode and cathode safety features on all components• Clear tank allows monitoring of sample progress• Buffer comes into contact with entire gel plate surface for even temperature control• Seamless moulded tank design with stirring bar corral• Compatible with pre-cast gels: CBS ClearPAGE™, Cambrex, and Invitrogen• Uses less than 1 litre of buffer for either application• No loose parts to keep track of on the central core

Applications:• SDS – PAGE

• Electroblotting (Western)

• Acrylamide nucleic acid separations


2 functions in 1! Mini-Vertical Slab Gel/Blotting System

The Dual Cool electrophoresis system

Get this unit FREE with a purchaseof ClearPAGETM gels and buffers

• Easy to use

• Versatile design

• Sturdy construction

See our promotion in

Shop!

>


For more information on these products contact your local VWR sales office,send an e-mail to [email protected] or visit our website www.vwr.comElectrophoresis

Additional features:• Available in 12, 17, 2D, and 1 well

formats, all 1,0 mm thick. High capacity wells in 2 and 17 wellformats give the same sensitivity as 1,5 mm thick 10 or 15 well gels.

• Gels are sold in packs of 10

• No comb or tape to remove shortensyour preparation time.

• Cassette design allows longer pathlength than competition, increasingresolution

• Well fingers project above the top of the cassette to prevent samplecarry-over from the outside of thepipette between lanes.

• Running buffers may be reused onthe anode side, unlike withNuPAGE™ gels. You can save 75% of the buffer cost.

• New precast gel technology with a unique acrylamide polymer forexceptional protein separations

• Ideal for western blots.

• ClearPAGE™ gels now have a reddye suspended within the loadingarea for better well visualisation.This dye runs ahead of thebromophenol blue marker.

ClearPAGE™ Precast Gels

For the best results,start with the world’s best precast gels

See our promotion in

Shop!Get a DCX-700 dual cool combo FREE witha purchase of ClearPAGETM gels and buffers

ClearPAGE™ offers superior quality, precast,composite polymer gels in 10 x 10 cm plasticcassettes for both SDS PAGE and DNA /NativePAGE. Recommended for use with C.B.S.Scientific’s DCX-700 Dual Cool Combo Cell.ClearPAGETM gels must use ClearPAGETM

Tris-Tricine buffers only. These buffers can eitherbe ordered or made using a ClearPAGETM bufferrecipe.

Long shelf life

Never throw away expired gels again! All ourgels use a neutral pH gel buffer to reduce alkaline hydrolysis. Combined with reducedpolymer swelling, all gels, including high percentage gels, carry a ”Best By Date” of overa year from the date of manufacture. Theyshow no change in the separation when storedat 4 ºC during that period, and will give usefulresults for years afterward. The gels also maybe stored at room temperature (up to 25 ºC)for 3 months after receipt.

Better resolution and sensitivity

The gel technology in ClearPAGE™ productsprovide robust separations superior to otherproducts on the market. The gel itself is longer(8 cm) and wider (8,5 cm) than competitors'gels to provide better separations andstraighter results. Improved stacking allowslarge sample volumes so that 1 mmClearPAGE™ cassette gels have the equivalentsensitivity to 1,5 mm gels. With theClearPAGE™ team members’ broad experiencedeveloping and making commercial pre-castgels and our commitment to quality, you can beassured of high-quality, consistent performanceevery time.

Superior DNA separations

Precast DNA and Native PAGE gels also have along shelf life. These gels are a neutral pH andutilise a stacking system. Larger sample volumes (such as 35 µl/12 well and 17 µl/17well) will still produce sharp bands.

Incredibly strong & elastic gels

New polymer and polymerisation technologyare used to make high-resolution, cross-linkedpolyacrylamide gels combined with a smallamount of agarose in plastic cassettes withouta coating. These gels are about 10 timesstronger than the equivalent polyacrylamide gelwithout agarose, so they can be handled,stained, and dried without tearing or cracking,and blotted without sticking to your blottingmembrane. This polymer also reduces shrinkingand swelling 10- to 20-fold allowing us tomake a full range of gel percentages using thesame gel and running buffer system. SDS PAGEseparations cover proteins from 1 to 1000 kDa.

Try ClearPAGE™ gels yourselfand find out why everyonesays they are the finest gelson the market.

For a full list of the gel types available orto request a FREE ClearPAGE sample kitcontact your local VWR sales organisationor email [email protected]

No sample loss or contamination,just high yields

Methods such as dialysis and electroelution (forextraction from gels) have long been used topurify proteins and nucleic acids. Thesetraditional methods may lead to sample loss andcontamination. D-Tube™ Dialyzers are a new,simpler way to dialyse sample volumes from 10-3000 µl. In addition, D-Tube™ Dialyzers canbe used for electroelution of protein, protein-protein or protein-DNA complexes,oligonucleotides, DNA, and RNA from 1D- and2D-polyacrylamide and agarose gels.

D-TubeTM Dialyzer features• Easy-to-handle dialyzers for buffer exchange

and removal of urea and detergents • One-step dialysis procedure that does not

require syringes or any special equipment • Efficient sample volume recovery (>97%)• Protease-, RNase-, and DNase-free • Ideal for electroelution of proteins, protein-

DNA complexes, oligo-nucleotides, DNA, andRNA from polyacrylamide and agarose gels

• Procedure compatible with variety ofdownstream applications including MALDI-MS, functional assays and HPLC

• Simultaneous electroelution from multiplesamples

D-Tube™ Dialyzers: maximisingconvenience and recovery

D-Tube™ Dialyzers are easy-to-handle dialysersin a capped centrifuge tube format with dialysismembrane windows for buffer exchange andremoval of solutes like urea or detergents whileproviding > 97% sample volume recovery. Thedisposable, single-use tubes require no syringes,centrifuge, or laborious steps to manipulatesmall sample volumes. The sample is added andremoved using a standard laboratory pipette.D-Tube™ Dialyzers are available with molecularweight cut-offs from 3,5 to 14 kDa and aredesigned with three volume capacities: mini(10–250 µl), midi (50–800 µl), and maxi(100–3000 µl).The membrane is ultra-clean, EDTA-treatedregenerated cellulose, sulphur- and heavymetal-free. Each kit contains 10 D-Tube™Dialyzers and one floating rack that can hold upto four devices in an exchange buffer.

Electroelution: recover proteinsand other biological moleculesfrom gelsFollowing separation of biomolecules byagarose or polyacrylamide gel electrophoresis,electroelution offers a means to extract thesample of interest by applying an electriccurrent to the excised gel band. Thecombination of D-Tube™ Dialyzers and the D-Tube™ Electroelution accessory kit providesa unique tool for extraction of any protein,protein-protein, or protein-DNA complexes fromnon-denaturing and denaturing (SDS)polyacrylamide gels, and for extraction ofoligonucleotides, RNA, and DNA from bothpolyacrylamide and agarose gels. The D-Tube™Electroelution accessory kit provides three D-Tube supporting trays that fit into mosthorizontal electrophoresis units and optimisedreagents for protein and nucleic acidprecipitation following electroelution.


D-Tube™ Dialyzers and D-Tube electroelution kit

Novel toolsfor samplepreparation


For more information on these products contact your local VWR sales office,send an e-mail to [email protected] or visit our website www.vwr.comSample preparation

Description Volume (µl) Pack Cat. No.

D-TubeTM Dialyzer Mini, MWCO 6–8 kDa 10 to 250 1 kit 71504-3 D-TubeTM Dialyzer Mini, MWCO 12–14 kDa 10 to 250 1 kit 71505-3 D-TubeTM Dialyzer Midi, MWCO 3,5 kDa 50 to 800 1 kit 71506-3 D-TubeTM Dialyzer Midi, MWCO 6–8 kDa 50 to 800 1 kit 71507-3 D-TubeTM Dialyzer Maxi, MWCO 3,5 KDa 100 to 3000 1 kit 71508-3 D-TubeTM Dialyzer Maxi, MWCO 6–8 KDa 100 to 3000 1 kit 71509-3 D-TubeTM Dialyzer Maxi, MWCO 12–14 KDa 100 to 3000 1 kit 71510-3

Description Pack Cat. No.

D-TubeTM Electroelution accessory kit 1 kit 71511-3

Description Volume (µl) Pack Cat. No.

D-Tube96TM Dialyzer Mini, MWCO 6–8 kDa 10 to 250 96 D-Tubes 71712-3 D-Tube96TM Dialyzer Mini, MWCO 12–14 kDa 10 to 250 96 D-Tubes 71713-3

Electroelution kit features • Efficient extraction of protein, protein-protein, and protein-DNA complexes, oligonucleotides,

DNA, and RNA from 1D- and 2D-polyacrylamide and agarose gels• More than 60% protein recovery in less than 2 hours• More than 90% recovery of oligonucleotides, RNA, and DNA of 15 nucleotides to 80 kbp• Procedure compatible with a variety of downstream applications including MALDI-MS,

functional assays and HPLC• Simultaneous electroelution from multiple samples

Components of each kit: • 10 D-TubeTM Dialyzers • 1 Floating rack

D-Tube™ Electroelution accessory kit

Components • 1 ml MS Precipitation Buffer • 10 ml TCA, 20%• 2 × 1 ml 3 M NaAc, pH 5,2• 3 supporting trays, Mini, Midi, Maxi

D-Tube96™ DialyzerConvenient and simultaneous dialysis of 96 samples in a high throughput manner

Components of each kit: • 1 D-Tube96TM Dialyzer (6-8 kDa or 12-14 kDa)(D-TubeTM Dialyzer Mini in a 96-tube configuration)

• 1 Aluminum Plate Sealer

Note: electroelution times shown in this table are for D-TubeTM Midi; optimum times depend on the samplecontents.For each D-TubeTM Midi, the gel piece should not exceed0,5 cm × 1 cm. Additional information about optimumelectroelution times for D-TubeTM Mini and Maxivarieties is available in TB422 at www.novagen.com

* Minimum time recommended for elution of protein froma 10% SDS-polyacrylamide gel at 100 V

† Minimum time recommended for elution of DNA orRNA fragments from a native or denaturing 4%polyacrylamide gel at 100–150 V

‡ Minimum time recommended for elution of DNAfragments from a 1% agarose gel at 80–110 V

Table 1:Typical recoveries using D-TubeTM Dialyzer Kits

Sample Type Method TypicalRecovery

Sample in solution Dialysis > 97%

DNA or RNA in agarose gel slice Electroelution > 90%

Oligonucleotides, DNA,or RNA in polyacrylamide Electroelution > 90%gel slice

Protein in polyacrylamide gel slice Electroelution 60%

Prot

ein

DNA

RNA

Olig

osDN

A

Table 2: Minimum electroelution times to extractsamples from polyacrylamide and agarose gels

POLYACRYLAMIDE GELSProtein Size (kDa) Elution Time* (min)

14 35–4519–26 45–55

29 55–6540 60–7045 65–7550 75–8566 85–9581 105–115116 120–130128 140–150

DNA Fragment Size (bp) Elution Time† (min)100 10–20300 15–25500 20–30822 25–35

1044 30–402700 45–55

RNA Fragment Size (nt) Elution Time† (min)100 15–25400 25–35600 35–45

1000 45–55Oligonucleotide Size (nt) Elution Time† (min)

15-100 10–20

AGAROSE GELSDNA Fragment Size (bp) Elution Time‡ (min)

100–200 10–20500–700 15–20

1000 20–304361 25–356557 45–559416 55–65

23130 70–80

The OpScanner is a simultaneous 2-colour scanner with confocal

PMT detection and a real-time autofocus system that combines

high resolution with low background noise and high sensitivity.

It’s data acquisition and image analysis software is easy to use

and like the unit itself robust and low maintenance. Taking up

little space on the laboratory bench the OpScanner is light-

weight and portable and designed to be right next to you when

you need it. It’s designed for individual lab use and has a very

affordable price tag.


The OpScanner is a reliable and easy-to-use scanner forall standard microarray slides

Microarray

Features

Simultaneous 2-colour scanner • Compatible with all standard microarray

slides • Fast scanning system: proprietary scanning

system allows less than 4 minutes per slidescan time

• High-resolution acquisition system: scansspot sizes from 200 µm to less than 30 µm

• Real-time image acquisition (accurate signaldetection for a variety of samples: DNA,protein microarrays, tissue or cell arrays)

Confocal PMT detection andreal-time autofocus system • Low background noise and high sensitivity • Uniform scanning across the whole biochip

surface whatever the substrate (glass andplastic)

• Not sensitive to shocks, vibrations and slidedeformation

Robust and easy-to-use software • Immediate and easy data acquisition and

image analysis software • Automatic spot recognition and powerful

image analysis • Software for various OS environments

(Linux, Windows) • Ethernet interface allows easy data sharing For more information contact your

local VWR sales organisation or [email protected]

Specifications

Optical system

Laser: Thermoelectrically cooled laser diode stabilised feedback output Excitation wavelength: 635 nm and 532 nm Compatible fluorophores: Cy5, Alexa 647, Alexa 660 (635 nm)

Cy3, Alexa 546, Alexa 555 (532 nm) Laser power: Adjustable (2 laser powers available) Gain PMT: Adjustable from 0 to 100% Autofocus: Real-time Detection: Real confocal and high performances digital PMT Optical resolution : 2,4 mm Pixel size: From 3 to 40 mm

Photometric performances

Dynamic range: Over 4 orders of magnitudeSensitivity: 0,1 fluor. / mm²Uniformity: < 5% CV

Interface & Software

Interface: EthernetImage format: TIFF (16 bits grey scale)Analysis image: Yes: accurate grid positioning, reliable calculations & export results

General

Sample: All standard microscope slide 25 x 75 mm / 1x3 "Scanning time: 3,5 min. per slideScanning speed: From 10 to 35 lines/sBarcode reader: All formatsPower supply: 100-240 VACDimension: 253 x 450 x 345 mm (L x D x H)

9,9 x 17,7 x 13,6 mm (L x D x H)Weight: 12 kg (26 lbs)



The analysis of drugs in hair was first introducedaround 1954. This method has the benefits ofpermitting a long-term retrospective recordingas well as allowing a relative quantification ofthe degree or intensity of drug use. As a result,this method has gained wide acceptance andhas been in use since the 1970s. To prepare ahair sample for analysis, the sample has to firstbe disrupted. This is most easily done using aball mill like the Mikro-Dismembrator S fromSartorius AG, Germany: the sample is defattedby washing in acetone and buffer, then dried,frozen and pulverized. The resultant cold powdercan be dissolved in a suitable buffer system. To

further simplify drug detection, the sample canthen be prepared using a Centrisart centrifugalultrafiltration device with a suitable molecularweight cut-off for removal of proteins containedin the sample.

The Mikro-Dismembrator ball mills are verypopular among scientists who have to

disrupt brittle or frozen material.With it's high performance at up to3.000 min-1 the new Mikro-Dismembrator S is particularlysuited for all applications whererapid and complete disruption ofthe samples is needed. Foroperation a shaking flask andsuitable grinding balls or glassbeads are required.

Centrisart 1 CentrifugalUnits are particularly suited forthis application because of theirspecial design. Centrisart 1consists of an outer sample tubeand an inner tube with anultrafilter bottom. The principle

of construction turns the traditional way offiltration upside down: ultrafiltration takes placein the opposite direction of the centrifugal force.This prevents blocking of the ultrafilter byconcentrate layers and even allowsultrafiltration of particle-containing samples likewhole blood or diluted homogenates. In mostapplications an ultrafilter with cut-off 20.000 issufficient for removal of proteins and othermacromolecules. The ultrafiltrate is collected inthe inner tube and can be easily removed forfurther testing.

Suggested further reading:- http://www.criminology.fsu.edu/journal/mcbay2.html -

a hair drug testing bibliography- P. Nebinger, M. Kroel:

J. Chromatography 619 (1993) 342-344- A. Gleixner, H. H. D. Meyer:

Fleischwirtschaft 76 (1996), 407 – 409 (incl. English references)

Sample preparation for drug testing in hair

Sample preparation

The use of illegal drugs is

connected to personal health

problems, social problems as well

as to potential safety risks in the

workplace or public traffic systems,

so the demand for tests is high.

In most cases, it is practically

impossible to determine the exact

point in time when the drugs were

consumed. As the human body

degrades the drugs, levels may

soon fall below detectable levels.

Hence, it is difficult to keep track

of the drugs when samples of

blood or urine are taken: even

though the samples will show

positive results when drugs have

been recently used, they will fail in

cases where use has been

abandoned or interrupted.

Isolation of high quality RNA is the first andoften most critical step in performing manyfundamental molecular biology experimentsincluding Northern analysis, nucleaseprotection assays, RT-PCR®, real-time RT-PCR®

and micro-array analysis. There are three majortechniques used extensively for RNAextraction: Organic extraction such as Phenol-Guanidine Isothiocyanate (GITC) basedsolution; Silica-membrane based spin columntechnology; Paramagnetic particles technology.One of the most commonly used methods isPhenol-Guanidine Isothiocyanate (GITC) basedorganic extraction, but RNA samples isolatedin this way are often contaminated withproteins and other cellular materials, organicsolvents such as phenol-chloroform, salts andethanol. Silica column and paramagneticparticles based RNA isolation systems do notrequire the use of toxic organic solvents, arerelatively simple and efficient, and yield totalintact RNA with low level contamination fromproteins and cellular materials. However, thosemethods can often result in significant level ofgenomic DNA contamination. Omega Bio-Tekhas developed diversified RNA isolation kitsthat cover almost all types of biological samplesources (see product list on page 38) withtechnologies that effectively reduce thegenomic DNA contamination. In this paper, wereport the test results from 4 commonly usedE.Z.N.A.™ RNA isolation kits to demonstratethe quality of RNA from different samplesources.

Materials and methodsSamples were collected from field or obtainedfrom commercial sources. Qitec Biotech Co. Ltdsupplied all the reagents except reagents fromkits.

RNA extractionRNA extractions were performed by followinguser instructions in each kit.

Real Time RT-PCR®

Real-time RT-PCR® analysis was performed usingthe Rotor-Gene 3000 (Corbett Life Science). 1 µgof purified RNA was reversed transcribed using 1 µg of random hexanucleotidic primers, 0,5 mMdNTP and 200 U M-MLV Reverse Transcriptase(Promega) at 37 °C for 1 hour in appropriatebuffer. SYBR Green Realtime PCR® Master Mix

(Toyobo) was used for real-time monitoring ofamplification according to the protocol. Thereal-time PCR® assay was carried out in avolume of 20 µl, 2 µM of each β-acitn primer,1 µl of template cDNA and Master Mix. Thermalcycling conditions were as follows: 95 °C for1min, 35 cycles of 95 °C for 15s and 55 °C for15s, 72 °C for 35s and 85 °C for 15s. Accurateamplification of the target amplicon waschecked by performing a melting curve.

ResultsWe have successfully isolated RNA from avariety samples using our four RNA isolationkits (E.Z.N.A.™ Total RNA Kit I, E.Z.N.A.™

Plant RNA kit, E.Z.N.A.™ Total RNA Kit II andE.Z.N.A.™ HP Total RNA Kit) and alsodemonstrated that the genomic DNAcontamination could be effectively eliminatedusing on-membrane DNase I digestion orE.Z.N.A.™ HP Total RNA protocol.

E.Z.N.A.™ Total RNA Kit I (R6834)The E.Z.N.A.™ Total RNA Kit use silica-membrane technology to isolate high quality ofRNA from 30 mg soft animal tissue or 1 x 107

Cells in 20 minutes. Briefly, culture cells andtissues are lysed and homogenised in TRK LysisBuffer (a highly denaturing guanidineisothiocyanate(GITC)-containing buffer).Ethanol is added to provide appropriate bindingconditions, and the sample is then applied toan HiBind RNA column. The column is thenwashed by Wash Buffer I and II. High-qualityRNA is then eluted in 50 µl of DEPC water.Figure 1 demonstrates that high quality RNAcan be successfully isolated from selectedsamples with typical yields shown in Table 1.Amplification results from real-time PCR® areshown in Figure 2.

E.Z.N.A.™ Plant RNA Kit (R6827)The E.Z.N.A.™ Plant RNA Kit is designed toisolate RNA from variety of plant species byusing silica-membrane technology. Figure 3demonstrates the high quality RNA successfullyisolated from various samples and the typicalyields are shown in Table 2.


High quality RNA isolation using E.Z.N.A.™ RNA systemTY, Zhu, Q, Guo

mg tissue A260 A280 Yield (µg)

Maize Leaves 100 0,5120 0,2561 40Tomato leaves 100 0,8750 0,4419 70Paddle leaves 100 0,3752 0,1923 30Maize seeds 100 0,2512 0,1275 20Paddle seeds 100 0,1625 0,0850 13

Table 2. Average yields of total RNA isolated from a varietyof plant using E.Z.N.A.™ Plant RNA kit.

Source No. cell mg tissue A260 A280 Yield (µg)

Cos-7 cell 5 x 106 0,8653 0,4323 70Hela cell 5 x 106 0,7532 0,3710 60LMH 5 x 106 0,6353 0,3100 50Mouse Liver 15 mg 0,7455 0,3826 60Mouse Spleen 15 mg 0,5368 0,2755 40

Table 1. Average yields of total RNA isolated from a varietyof cells and tissues using E.Z.N.A.™ Total RNA Kit

Figure 1. 1% agarose gel analysis of totalRNA isolated with E.Z.N.A.™ Total RNAKit I from Cos-7 cells (1), Hela cells (2),LMH cells (3), Mouse Liver (4) and MouseSpeed (5). 5% of purified RNA wereloaded on gel.

E.Z.N.A.™ Total RNA Kit II (R6934)

The E.Z.N.A. Total RNA Kit II takes theadvantage of one step RNA isolationtechnology and silica-membrane technologyand combines these two systems together.Although this kit is mainly designed for fattytissues, it can be used for almost all types ofbiological samples. With two extraction steps,this kit can significantly reduce the level ofgenomic DNA contamination (Figure 4) andincrease the purity of RNA. Figure 5demonstrates result from real-time RT-PCR®.

E.Z.N.A.™ HP Total RNA Kit(R6812)

The E.Z.N.A.™ HP Total RNA Kit is designed forisolating RNA from small amounts of cells andtissues. By using a specially designed genomicDNA removal spin column, this kit effectivelyremoves genomic DNA. To show the efficacy ofthis kit, Mouse Liver, Mouse Spleen and Cos-7cells were used isolate RNA with three kits:E.Z.N.A.TM Total RNA Kit I, E.Z.N.A.TM Total RNAKit II and E.Z.N.A.TM HP Total RNA Kit. AfterPurification, the RNA were treated with RNaseA (No DNase I detected) and then extracted byphenol, precipitated by isopropanol, and anyDNA re-dissolved by Buffer TE. DNAcontamination level using different kits isshown in Figure 6.

On-Membrane DNase I digestion(E1091)

Omega Bio-tek has developed an on-membraneDNase I protocol that can be use in allE.Z.N.A.™ RNA Kits to remove the genomicDNA during RNA isolation process. To show theefficiency of on-membrane DNase I digestion,the RNA and DNA were bound to a column,then treated with DNase I (E1091) as per kitinstructions, the results were showed in Figure7 and Figure 8.


For more information on these products contact your local VWR sales office,send an e-mail to [email protected] or visit our website www.vwr.comRNA extraction

Figure 2. RNA waspurified from Hela cells,Cos-7 cells and 293 cellusing the E.Z.N.A.™Total RNA Kit I (treatedwith DNase I). Afterreverse transcription,real-time, quantitativePCR® was carried out onthe Rotor-Gene 3000and PCR® specific for β-actin.

Figure 4. 1% agarose gel analysis ofRNA purified with E.Z.N.A.TM Total RNAKit II from pig heart (1), liver (2), kidney(3), fat (4), spleen (5) and brain (6). 1% ofpurified RNA was loaded on gel.

Figure 6. RNA was isolated from differentsamples with three kits. And the DNA contaminants are showed as figure.

Figure 5. RNA was purifiedfrom pig fatty tissue using theE.Z.N.A.TM Total RNA Kit II. After reverse transcriptionusing M-MLV ReverseTranscriptase, real-time,quantitative PCR® was carriedout on the Rotor-Gene 3000and primers specific for β-actin.

Figure 8. RNA treated with (D1) or without (1) DNase I on membrane, thenPCR® ampilified β-actin.

Figure 7. Effect of On-Membrane DNase Idigestion treatment: Total RNA isolated withE.Z.N.A.™ Total RNA Kit I from Cos-7 cells.Then the column is treated with DNase I (D1)or without treated (1). RNA eluted withDEPC-Treated water. 10% of eluted RNA wasanalysed by 1% agarose gel.

Figure 3. 1% agarose gel analysis of totalRNA isolated from plant with two protocolsfrom E.Z.N.A.™ Plant RNA Kit: Standard protocol: 1: Paddle leaves, 2: lichi leaves, 3: Tomato leaves, 4: MaizeLeaves: Optional protocol for difficult samples: 5: Paddle seeds, 6: Maize seeds, 7: Potato leaves, 8: pumpkin leaves.

D1 D1 1 1

Conclusion and discussionThe E.Z.N.A.™ RNA range includes specialisedkits for different types of starting materialsinclude all types of animal tissues, bacteria,plant, fungi, yeast and molluscs. Different kitformats are available handling small and largesample sizes and for manual and automatedprocessing of up to 96 or 192 samples inparallel. A previous study showed that noexisting RNA isolation method can completelyavoid the genomic DNA contamination(Vongsavanh, 2002). Omega Bio-Tek hasdeveloped two methods to significantly reducethe genomic DNA contamination. The E.Z.N.A.TM

RNA purification kits, which are based on silica-membranes and Mag-BindTM RNA purificationkits provide a complete solution for consistentyield of high quality RNA.

Omega Bio-tek has developed a full range ofRNA isolation kits to provide reliable RNApurification procedures based on the uniqueproperties of biological samples. For example,when purifying RNA from animal tissue, somesamples such as liver can be easily lysed withlysis buffer, while other samples such as braintissue or muscle tissues require specialtreatments for lysis. Plant tissues containdifferent levels of phenolic compounds and/orpolysaccharides. The E.Z.N.A.™ RNAPurification kits, which are all based on silica-membrane technology except Mag-Bind™ RNApurification kits, provide a completely solutionfor consistent yield of highly pure RNA. Visitwww.omegabiotek.com to find out more aboutnew solutions for RNA purification.


Samples Product Size Cat. No.

Soft animal tissue: 30 mg Total RNA Kit I 50 OMEGR6834-01Culture cells: 1 x 107 cells 200 OMEGR6834-02Soft animal tissue: 100 mg Total RNA Midi Kit 10 OMEGR6664-01Culture cells: 1 x 108 cells 25 OMEGR6664-02Soft animal tissue: 1g Total RNA Maxi Kit 5 OMEGR6693-01Culture cells: 5 x 108 cells 20 OMEGR6693-02100 mg tissue or 1 x 107 culture cells Total RNA Kit II 50 OMEGR6934-01

200 OMEGR6934-02Fibrous or fixed tissue: 30 mg Tissue RNA Kit 50 OMEGR6688-01

200 OMEGR6688-02Tissue: <10 mg, MicroElute RNA Kit 50 OMEGR6831-01Cells: < 1 x 106 cells 200 OMEGR6831-021 ml blood Blood RNA Kit 50 OMEGR6814-01

200 OMEGR6814-0210 ml blood Blood RNA Midi Kit 10 OMEGR6615-01

25 OMEGR6615-0250 ml blood Blood RNA Maxi Kit 5 OMEGR6616-01

20 OMEGR6616-02100 mg plant Plant RNA Kit 50 OMEGR6827-01

200 OMEGR6827-02500 mg plant Plant RNA Midi Kit 10 OMEGR6628-01

25 OMEGR6628-025 g plant Plant RNA Maxi Kit 5 OMEGR6629-01

20 OMEGR6629-02100 mg fungal Fungal RNA Kit 50 OMEGR6840-01

200 OMEGR6840-021-3 ml log phase bacterial Bacterial RNA Kit 50 OMEGR6950-01

200 OMEGR6950-021-3 ml log phase yeast Yeast RNA Kit 50 OMEGR6870-01

200 OMEGR6870-02Serum and cell free body fluids Viral RNA Kit 50 OMEGR6874-01

200 OMEGR6874-02Cell and tissues DNA/RNA Kit 50 OMEGR6731-01

200 OMEGR6731-02Soft animal tissue: 30 mg E-Z 96 Total RNA Kit 4 x 96 OMEGR1034-01Culture cells: 1 x 107 cells 12 x 96 OMEGR1034-02Fibrous or fixed tissue: 30 mg E-Z 96 Tissue RNA Kit 2 x 96 OMEGR1088-01

8 x 96 OMEGR1088-02Serum and cell free body fluids E-Z 96 Viral RNA Kit 4 x 96 OMEGR1074-01

High quality RNA isolation using E.Z.N.A.™ RNA system(continued)

All you need for PCRAll you need for PCR®®

This handy brochure is an essential tool for molecular biologists covering instrumentation and reagents. Order your copy from your local VWR sales organisation or email [email protected]


For more information on these products contact your local VWR sales office,send an e-mail to [email protected] or visit our website www.vwr.comCell manipulation

Control the power – transfer the genesBTX New Electro Cell Manipulator – the perfect tool forfusion, nuclear transfer and electroporation applications

Model Cat. No.

ECM 2001 Embryo Manipulation System 732-0615ECM 2001 Electro cell Fusion System 732-0616ECM 2001 Electroporation System 732-0802ECM 2001 Generator only 732-0617

BTX/Harvard Apparatus, a leader inelectroporation and electrofusion instrumentsand accessories, offers the ECM 2001 ElectroCell Manipulator (ETL and CE marked). Thisinstrument is a safe, user-friendly tool for genetransfer, nuclear transfer and cell fusion of avariety of plant, animal and bacterial cells.

The advanced functions of the ECM2001 areideal for in vivo and in vitro experimentsresulting in hybridomas, transfected cancercells, and engineered embryonic stem cells(ESC), and for in vivo drug delivery andexperimental cancer therapy. Nuclear transferapplications until now resulted in low efficiencyand low viability. The novel techniqueemploying electrofusion to nuclear-transfercomplexes improves both efficiency andviability. In short oocyte-cell couplets arealigned in a BTX fusion chamber by applying AC

pulse. The fusion of the couplets isaccomplished by applying two DC pulses, and apost-fusion AC pulse.

A variety of electrodes incorporated in cuvettesand microslides are available for single cellexperimentation, and genetrodes,tweezertrodes and genepaddles for in vivo testson tissues and organs. The instrument’s CEmark and ETL mark allows it to be a truly globalinstrument applicable for experimental workaround the world. The superior design carries atwo year warranty from the day of purchase.BTX’s global technical support team will assistscientists worldwide to optimise theirapplications and choose the best accessoriesfor their applications.

• AC feature aligns cells and maintains cell membrane compression

• Begin fusion cell and membranesusing the DC square wave pulse

• Cell fusion complete!

Cell fusion

ABsoluteTM Blue is a range of high performance master mixes specially formulated to achieve themost consistent and reproducible QPCR data across all QPCR platforms. Like ABgene’s ABsoluteTM

Mixes, proprietary additives give enhanced and consistent endpoint readings as well as low Ctvalues. All contain ABgene’s Thermo-Start® hot-start enzyme, for increased sensitivity, and replacedUTP with dTTP for increased reaction efficiency and reduced time per reaction (low Ct).

Uniquely, ABsoluteTM Blue incorporates an inert dye to significantly enhance the contrast betweenreagent and plastic, making verification of master mix dispensing quick, easy and foolproof. This isparticularly important for users of opaque white plates and tubes. Although recognised as givingsuperior QPCR results, traditional colourless reagents are harder to visualise. With ABsoluteTM Blue, asimple glance ensures that the correct amount of master mix has been added.

ABsoluteTM Blue Master Mixes are ideal for QPCR analysis in molecular biology, including geneexpression and SNP typing.

ABsoluteTM Blue – clearly superiorCompare ABsoluteTM Blue QPCR Master Mix with the competition and a clear winner emerges.

Now, you can achieve similar consistency and reproducibility while minimising the risk ofaliquotting errors. Switch to ABsoluteTM Blue and transform ease-of-use for your QPCR.

Transforms ease-of-use

and yields significant

performance advantages

for all QPCR users


ABsolute™ Blue - the advantage is clear

>ABsoluteTM Blue - clearly better

• High performance – ABsolute™ sensitivity, consistency & convenience

• Enhanced contrast and reduced errors – easily identify correctly aliquottedreaction mix in all plates and tubes

• Universal – compatible with all QPCRplatforms

• Flexible – use with probes or as SYBRGreen mix

• Adaptable – available with ROX orFluorescein passive reference dyes

• Thermo-Start® – increased sensitivity

• dTTP – increased PCR® reaction efficiencyand lower Ct values

• Licensed for QPCR

Figure 1 shows a iQ BioRad QPCR cycler amplification/cycle graph of a 295bptarget of the ß-Actin gene via SYBR®

Green chemistry using a ten-fold dilution series of human genomic DNA(200 ng-200 pg) in triplicate reactionscomparing ABsolute™ Blue QPCRMaster Mix (Blue) and CompetitorSYBR Green QPCR Master Mix (Green).

See our special offer on white platesin the VWRbioMarke shop issue 7 Shop!


For more information on these products contact your local VWR sales office,send an e-mail to [email protected] or visit our website www.vwr.comQPCR

Description Pack Size (number of rxns)200 x 25 µl 1600 x 25 µl 400 x 25 µl 4000 x 25 µl 4000 x 25 µl 40000 x 25 µl 2 x 1,25 ml 16 x 1,25 ml 1 x 5 ml 10 x 5 ml vial 1 x 50 ml vial 10 x 50 ml

ABsoluteTM Blue QPCR Mix plus ROX vial 733-0942 733-0943 733-0944 733-0945 733-0946 733-0947ABsoluteTM Blue QPCR ROX Mix 733-0948 733-0949 733-0950 733-0951 733-0952 733-0953ABsoluteTM Blue QPCR Low ROX Mix 733-0972 733-0973 733-0974 733-0975 733-0976 733-0977ABsoluteTM Blue QPCR SYBR® Green Mix plus ROX vial 733-0960 733-0961 733-0962 733-0963 733-0964 733-0965ABsoluteTM Blue QPCR SYBR® Green ROX Mix 733-0954 733-0955 733-0956 733-0957 733-0958 733-0959ABsoluteTM Blue QPCR SYBR® Green Fluorescein Mix 733-0966 733-0967 733-0968 733-0969 733-0970 733-0971ABsoluteTM Blue QPCR SYBR® Green Low ROX vial 733-0978 733-0979 733-0980 733-0981 733-0982 733-0983

ABsoluteTM Blue – clearly visible

One of the issues facing QPCR users is ensuring that the correct amount of master mix is added tothe reaction. Since most QPCR master mixes are clear, it is often difficult to discern which wellshave been aliquotted successfully, especially when using white plates.

ABsoluteTM Blue QPCR Master Mixes overcome this problem. They use an inert blue dye, which doesnot effect the QPCR reaction, to significantly enhance the contrast between reagent and plastic. Theblue colour offers an immediate check that a well has not been missed when dispensing manually.Also, a scan across the strip or plate will identify any differences in colour intensity that mightindicate inaccurate dispensing. This is an especially useful and rapid validity check when using anautomated dispensing system, where variation in colour may indicate a blockage in one or more ofthe dispensing channels.

dTTP vs. dUTP – clearly desirableAlmost all currently available master mixes use dUTP in conjunction with Uracil-N-Glycosylase(UNG) to remove any amplicon carry-over contamination from previous reactions by degradingthose templates containing dUTP. However, dUTP forces the Taq polymerase to move more slowlyalong the template, which can increase incorporation mistakes and decrease the overall efficiencyof the reaction.

ABgene insists on the use of dTTP in its master mixes. A naturally occurring base, dTTP is morereadily incorporated into the amplicon. Its incorporation into the master mixes increases thereaction efficiency by approx. 5% and reduces the reaction time.

White consumables – clearly sensitiveThe use of clear plastic consumables allows the fluorescent signal to pass through the well wall andhit the Peltier block. Inconsistencies in the Peltier block reduce the reflected signal, which is alsorefracted as it passes back through the clear plastic. This means that variations between replicatesmay be due to this inefficient reflection/refraction of the signal rather than any true differencesbetween the samples.

ABgene opaque white polypropylene plastics reflect the fluorescent signal back to the detectormore efficiently than clear plastics. More efficiently, in fact, than any other colour – including whiteplastic from other manufacturers. The result is significantly improved sensitivity and consistencybetween wells.

GAPDH amplification using 2 ngHuman genomic DNA SYBR® Green Mixin Natural plates (Blue) and in whiteplates (Red).

ABgene* products are not available in Italy,Belgium and the Netherlands

For a free sample contact [email protected] stating the volume you require.


For more information on these products contact your local VWR sales office,send an e-mail to [email protected] or visit our website www.vwr.comLiquid handling

The Low Retention Tip on the left has virtually no sample left behind, in contrast to the standardtip on the right.

Continuing our mission to provide the scientific community with pipette

tips of the highest quality, Molecular BioProducts Inc. highlights

Low Retention pipette tips for greater accuracy and no contamination.

Low Retention technology is based on a proprietary process that makes our pipette tips morehydrophobic, allowing researchers to load dyes, surfactants, and solvents and to pipette viscoussamples much more accurately then ever before, simply because there is much less sample retainedwithin the tip.

Low Retention Tips

Description Volume Pack Cat. No.

ART 10 10 µl 96 tips/tray, 10 trays/pack 732-2220ART 10 REACH 10 µl extended length 96 tips/tray, 10 trays/pack 732-2222ART 10F 10 µl 96 tips/tray, 10 trays/pack 732-2286ART 20E 10 µl 96 tips/tray, 10 trays/pack 732-2294ART 20P 20 µl 96 tips/tray, 10 trays/pack 732-2224ART 100E 100 µl 96 tips/tray, 10 trays/pack 732-2205ART 200 200 µl 96 tips/tray, 10 trays/pack 732-2208ART 1000 1000 µl 100 tips/tray, 8 trays/pack 732-0995ART 20 SoftFit 20 µl 96 tips/tray, 10 trays/pack 732-2314ART 200 SoftFit 200 µl 96 tips/tray, 10 trays/pack 732-2316ART 1000 SoftFit 1000 µl 100 tips/tray, 8 trays/pack 732-2318

See for our very special offer in Shop!

Top selling Low Retention ART barrier tips

These extremely hydrophobic tips are available with the ART® self-sealing barrier, PureTM, which areboth independently certified to be RNase-, DNase-, DNA- and pyrogens-free - Crystal-clear, qualityracked MBP® tips with the MicropointTM design without filters are also available.

Your European Distribution Partner

AustriaVWR International GmbHGraumanngasse 71150 WienTel.: 01 97 002 0Fax: 01 97 002 600E-mail: [email protected]

BelgiumVWR International bvba/sprlHaasrode Researchpark Zone 3Geldenaaksebaan 4643001 LeuvenTel.: 016 385 011Fax: 016 385 385E-mail: [email protected]

DenmarkVWR International ApSValhøjs Alle 174-1762610 RødovreTel.: 43 86 87 88Fax: 43 86 87 90E-mail: [email protected]

FinlandVWR International OyPihatörmä 1 C 102240 EspooTel.: 09 80 45 51Fax: 09 80 45 52 00E-mail: [email protected]

FranceVWR International S.A.S.Le Périgares – Bâtiment B201, rue Carnot94126 Fontenay-sous-Bois cedexTel.: 0 825 02 30 30 (0,15 EUR TTC/min)

Fax: 0 825 02 30 35 (0,15 EUR TTC/min)

E-mail: [email protected]

GermanyVWR International GmbHHilpertstrasse 20aD - 64295 DarmstadtTel.: 0180 570 20 00Fax: 0180 570 22 22E-mail: [email protected]

IrelandAGB Scientific Ltd - A VWR International CompanyOrion Business CampusNorthwest Business ParkBallycoolinDublin 15Tel: 01 88 22 222Fax: 01 88 22 333E-mail [email protected]

ItalyVWR International s.r.l.Via Stephenson 9420157 Milano (MI)Tel.: 02 332 03 11Fax: 800 152 999E-mail: [email protected]

The NetherlandsVWR International B.V.Postbus 81981005 AD AmsterdamTel.: 020 - 4808 400Fax: 020 - 4808 480E-mail: [email protected]

Northern IrelandAGB Scientific Apparatus LtdA VWR International CompanyA10 Harbour Court7 Heron RoadSydenham Business ParkBT3 9HB BelfastTel.: 028 9058 5800Fax: 028 9080 7812E-mail: [email protected]

NorwayVWR International ASKakkelovnskroken 1P.B. 45, Kalbakken 0901 OsloTel.: 02290Fax: 22 90 00 40E-mail: [email protected]

PortugalVWR International – Material deLaboratorio, Lda. PortugalRua Alfredo da Silva 3-C1300-040 LisboaTel: 21 3600 770Fax: 21 3600 798/9E-mail: [email protected]

SpainVWR International Eurolab S.L.Apartado 4808100 Mollet del Vallés - BarcelonaTel.: 902 222 897Fax: 902 430 657E-mail: [email protected]

SwedenVWR International ABFagerstagatan 18a163 94 StockholmTel.: 08 621 34 00Fax: 08 621 34 66E-mail: [email protected]

SwitzerlandVWR International AGLerzenstrasse 16/18 8953 DietikonTel.: 044 745 13 13Fax: 044 745 13 10E-mail: [email protected]

UKVWR International LtdCustomer Service CentreHunter BoulevardMagna ParkLutterworthLeicestershireLE17 4XNTel.: 0800 22 33 44Fax: 01455 55 85 86E-mail: [email protected]

Standardize microarrays with automation

• New standards in microarray processing• Bubble free hybridization (unique ABSTM System)• Reproducible and reliable results• Multi-segment formats• Shortened process times• Unmatched flexibility

Tecan’s HS ProTM series & Tecan LS ReloadedTM

Liquid Handling & Robotics � Detection � Sample Management � Components � Services & Consumables

To find out more about our microarray products, please contact your local VWR Representative for more information. www.vwrbiosciences.com

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