For Research Use Only - Not for use in diagnostic procedures.

SUPERNOVA POLYMER DYESA STELLAR NEW WAY TO SEE DIM POPULATIONS

For Research Use Only - Not for use in diagnostic procedures.

INTRODUCING THE NEXT GENERATION OF POLYMER DYES WITH IMPROVED STAINING INDEX: SUPERNOVA V428

Beckman Coulter Life Sciences introduces the next generation of polymer dyes to bring cutting edge science to clinical research.

SuperNova conjugated antibodies not only deliver unique brightness for flow cytometry staining, but also generate limited nonspecific staining thanks to a proprietary formulation. Figure 1: Absorption and emission spectra (lexc = 405 nm

of SuperNova v428 in PBS).

SuperNova v428 (SN v428), the first dye of the series, is optimally excited by the violet laser (405 nm) with an excitation maximum of 414 nm. It has an emission peak of 428 nm and can be detected using a 450/50 bandpass filter or equivalent. It is among the brightest dyes excitable by the violet laser, and is therefore particularly suited for the assessment of dimly expressed markers.

As shown in Table 1 and Figure 2, SuperNova v428 conjugated antibodies deliver brighter signal compared to their BV421 equivalent, and consequently greater discrimination between negative and positive populations, making them well-suited to stain dim populations. SN v428 conjugates also generate significantly less non-specific staining to help ensure more confidence in the results. SN v428 conjugates are a brighter alternative to Pacific Blue*, being on average more than 10 times brighter, and the only fluorochrome excitable by the violet laser delivering a brightness even superior to PE conjugates.

SN v428 can be provided conjugated to antibodies through LUCID Custom Design Services.

Lymphocytes Stain Index

Monocytes MFI (background)

CD19

SN v428 281 0,688BV421 191 1,533PE 246 0,805Pacific Blue* 48 0,600

CD22

SN v428 172 1,033

BV421 123 1,476

PE 136 0,716

Pacific Blue* 28 0,958

CD25

SN v428 36 0,731

BV421 14 2,021

PE 24 1,052

Pacific Blue* 6 0,522

Table 1: Lymphocytes staining index and monocytes MFI (background) obtained staining normal whole blood with lymphocytes markers conjugated to SN v428, BV421, PE and Pacific Blue*.

Accelerating AnswersFor Research Use Only - Not for use in diagnostic procedures.

FLOW CYTOMETRY RESULTS

Figure 2: Staining pattern comparison of SN v428, BV 421, PE and Pacific Blue* conjugated to CD19, CD22, and CD25. SN v428 systematically provided highest brightness. Non-specific staining is systematically reduced compared to BV 421 conjugates.

Sup

erN

ova

v428

Bri

llian

t V

iole

t 4

21P

EP

acifi

c B

lue*

CD19 CD22 CD25

[WBC] FL9 INT / SS INT [WBC] [WBC] FL9 INT / SS INT

[WBC] FL9 INT / SS INT [WBC] [WBC] FL9 INT / SS INT

[WBC] FL2 INT / SS INT [WBC] [WBC] FL2 INT / SS INT

[WBC] FL9 INT / SS INT [WBC] [WBC] FL9 INT / SS INT

FL9 INT

FL2 INT FL2 INT FL2 INT

FL9 INTFL9 INTFL9 INT

FL9 INTFL9 INTFL9 INT

SS IN

T

SS IN

T

SS IN

T

SS IN

T

SS IN

T

SS IN

TSS

INT

SS IN

T

SS IN

TSS

INT

SS IN

T

SS IN

T

FL9 INT FL9 INT

For Research Use Only - Not for use in diagnostic procedures.

SUPERNOVA POLYMER DYES TECHNOLOGY

+Monomers

A B1- Polymerization

2- COOH capping

hv hv hv hv hv hv hv

1. Polymers are made from different monomers

2. Each monomer can absorb light and reemit at longer wavelength

3. Tandems can be derived from core polymers

4. Both core and tandem polymers can be conjugated to antibodies

Acceptor dye

ep or dcceAAccAcAcA c

dddyyeeyeydy

Tandems can be derived from core polymers

Acceptor dye

hv hv hv hv hv hv hv

Both core and tandem polymers can be conjugated to antibodies Both core and tandem polymers can be conjugated to antibodies Both core and tandem polymers can be conjugated to antibodies

Accelerating AnswersFor Research Use Only - Not for use in diagnostic procedures.

SuperNova v605 and SuperNova v786 are tandem polymer dyes, derived from the core SN v428. Therefore both share the same absorbance characteristics, with a maximum excitation at 414 nm. With emission peaks respectively at 605 and 786nm, they are optimally detected using the 610/20 and 780/60 nm bandpass filters of the flow cytometer.

Figure 3: Absorption and emission spectra of SN v605 and 786 (lexc = 405 nm of SuperNova v428 in PBS).

SuperNova v605 conjugated to CD20 and CD38, and SuperNova v786 conjugated to CD200 and HLA-DR antibodies were compared to their BV610 and BV786 equivalent on normal whole blood samples. As shown in table 2 and Figure 4, the SuperNova conjugated Antibodies deliver brighter signal compared to their BV dyes counterpart, and consequently greater discrimination between negative and positive populations. SN v605 and SN786 conjugates also generate significantly less non-specific staining.

SN v605 and SN786 can be provided conjugated to antibodies through LUCID Custom Design Services.

Lymphocytes Stain

IndexMonocytes MFI (background)

CD20SNv605 660.61 1101.65

BV605 522.62 1108.40

CD38SNv605 11.51

N/A§

BV605 7.12

CD200SNv786 16.16 2821.94

BV786 7.17 4609.08

HLA-DRSNv786 46.01

N/A§

BV786 26.37

Table 2: Lymphocytes staining index and monocytes MFI (background) obtained staining normal whole blood with lymphocytes markers conjugated to SN v605, SN v786, BV605 and BV786.

SUPERNOVA V605 AND SUPERNOVA V786

§ CD38 and HLA-DR staining on monocytes is not background, both being naturally expressed on monocytes

For Beckman Coulter’s worldwide o¢ce locations and phone numbers, please visit “Contact Us” at beckman.com/home

20.12.386.FLOW

© 2020 Beckman Coulter, Inc. All rights reserved. Beckman Coulter, the stylized logo, and the Beckman Coulter product and service marks mentioned herein are trademarks or registered trademarks of Beckman Coulter, Inc. in the United States and other countries.

* Pacific Blue is a registered trademark of Molecular Probes Inc.

For Research Use Only - Not for use in diagnostic procedures.

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FLOW CYTOMETRY RESULTS

CD

200

HLA

-DR

SuperNova v786Brilliant Violet 786

[WBC] Violet780-A / SSC-A[WBC] Violet780-A / SSC-A

[WBC] Violet780-A / SSC-A[WBC] Violet780-A / SSC-A

Violet780-AViolet780-A

Violet780-A Violet780-A

SSC

-ASS

C-A

SSC

-ASS

C-A

SuperNova v605Brilliant Violet 605

CD

38C

D20

[WBC] Violet610-A / SSC-A

SSC

-A

Violet610-A Violet610-A

SSC

-A

[WBC] Violet610-A / SSC-A

[WBC] Violet610-A / SSC-A [WBC] Violet610-H / SSC-A

SSC

-A

SSC

-A

Violet610-HViolet610-A