8/6/2019 ChromoTek RFP Booster

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Chromotek-RFP-Trap Booster

Product List

Chromotek-RFP-Trap Booster ACT-CM-RBOOSTR 100 ug

Allele Biotech-Introducing Cost Effectiveness to Research

F or Research Use Only. Not for Diagnostic or Therapeutic Use.Purchase does not include or carryany right to resell or transfer this prod-uct either as a stand-alone product or as a component of another product.

Any use of this product other thanthe permitted use without the expresswritten authorization of Allele Biotechis strictly prohibited

Website: www.allelebiotech.comCall: 1-800-991-RNAi/858-587-6645(Pacic Time: 9:00AM~5:00PM)Email: oligo@allelebiotech.com

For Technical Support:Email:FP@allelebiotech.com

Description RFP-T rap coupled to uorescent dye ATTO 594

Specicity RFP-Trap efciently highlights monomeric derivates of dsRed(mRFP1, mCherry,mOrange, mPlum), but not GFP and itsvariants.

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Relevance Red fluorescent proteins (RFPs) are widely used as fluorescent tags.Commonly, a combination of red and green fusion proteins (GFPs) isemployed to allow multicolor tracking and analysis of protein-proteininteractions. In most applications, the ability to observe fluorescence fromred florescent proteins is hindered because the quantum yield and photo-stability of RFPs are usually not sufficient enough for their microscopicdetection. This limitation is particularly important for Super ResolutionMicroscopy applications (e.g. 3D-SIM or STED). Many cell biologicalmethods such as HCl treatment for BrdU-detection, the EdU-Click-iTtreatment or heat denaturation for FISH, lead to disruption of the RFPsignal. By using RFP-Booster_Atto594, a specific RFP-binding proteincoupled to the fluorescent dye ATTO 594 (from ATTO-TEC), your RFPsignal will significantly increase in stability and brightness.

ATTO 594 is a fluorescent label belonging to the class of Rhodamine dyes.Fluorescence is excited most efficiently in the range 560 - 615 nm (abs= 601nm), a standard 561 nm laser is also suitable for excitation. Maximum fluore-scence emission falls in the range 615 - 680 nm (fl= 627 nm).

For further information please refer to http://www.atto-tec.com

Tested Appli-cations

Immunouorescence

Conc. 1 mg/ml

Storage Shipped at RT. Store at 4C Avoid freez ing

1. Fixation: 4% paraformaldehyde(PFA) or 1:10 formalin (37% formal-dehyde, 10-15% MetOH) in PBS, 10min., RT.

2. Wash 3x with PBS containing0.1% Tween 20 (PBST). Critical: donot let coverslips dry.

3. Permeabilisation: PBS contain-ing 0.5% Triton X-100, 5 min., RT.

Alternatively permeabilise by incu-bating in 100% methanol for 5min

at -20C.

4. Wash 2x with PBST.

5. Blocking: 4% BSA in PBST, 10min, RT.

6. RFP-Booster incubation: diluteRFP-Booster 1:200-1:400 in blockingbuffer and incubate 1 h, RT. *7. Wash 3x 5-10 min in PBST.

8. If required counterstain with DNAuorescent dyes, e.g. DAPI.

9. Before mounting, briey rince coverslipsbe very rinsed in water to prevent salt crystal formation.

10. Mount in VectaShield (Vector Labs) or other mounting media withanti-fading agents and seal mountedcoverslips with clear nail polish.

Please note: Optimal dilutions/ concentrations

should be determined by the end user.

*Tip: RFP-Booster can be added to master-mixduring routine immunostaining, along with theprimary or secondary antibodies.

Protocols

HeLa cells expressing mRFP-PCNA fixedand permeabilized. Staining with RFP-Booster_Atto594 (dilution 1/200, 1 h at RT).