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Brilliant Violet 510™ anti-mouse/human CD45R/B220 Antibody

Pricing & Availability
Clone
RA3-6B2 (See other available formats)
Regulatory Status
RUO
Other Names
B220
Isotype
Rat IgG2a, κ
Ave. Rating
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Product Citations
publications
1_RA3-6B2_BV510_CD45R_Antibody_1_FC_051816
C57BL/6 mouse splenocytes were stained with CD45R/B220 (clone RA3-6B2) Brilliant Violet 510™.
  • 1_RA3-6B2_BV510_CD45R_Antibody_1_FC_051816
    C57BL/6 mouse splenocytes were stained with CD45R/B220 (clone RA3-6B2) Brilliant Violet 510™.
  • 2_RA3-6B2_BV510_CD45R_Antibody_2_IHCF_051816
    C57BL/6 mouse frozen spleen section was fixed with 4% paraformaldehyde (PFA) for ten minutes at room temperature and blocked with 5% FBS plus 5% rat/mouse serum for 30 minutes at room temperature. Then the section was stained with 1 µg/ml of anti-mouse/human CD45R/B220 (clone RA3-6B2) Brilliant Violet 510™ (green), anti-mouse CD8a (clone 53-6.7) Brilliant Violet 421™ (blue) and anti-mouse Ly-6G (clone 1A8) Alexa Fluor® 647 (red) overnight at °C. The image was captured with a 10X objective.
  • 3_50_Mouse_Gut_EpCAM_CD117_B220
    Confocal image of C57BL/6 mouse small intestine sample acquired using the IBEX method of highly multiplexed antibody-based imaging: EpCAM (cyan) in Cycle 1, B220 (purple) in Cycle 2, and CD117 (yellow) in Cycle 2. Tissues were prepared using ~1% (vol/vol) formaldehyde and a detergent. Following fixation, samples are immersed in 30% (wt/vol) sucrose for cryoprotection. Images are courtesy of Drs. Andrea J. Radtke and Ronald N. Germain of the Center for Advanced Tissue Imaging (CAT-I) in the National Institute of Allergy and Infectious Diseases (NIAID, NIH).
Compare all formats See Brilliant Violet 510™ spectral data See high resolution IHC data...
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103247 125 µL 132€
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103248 50 µg 194€
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Description

CD45R, also known as B220, is an isoform of CD45. It is a member of the protein tyrosine phosphatase (PTP) family with a molecular weight of approximately 180-240 kD. CD45R is expressed on B cells (at all developmental stages from pro-B cells through mature B cells), activated B cells, and subsets of T and NK cells. CD45R (B220) is also expressed on a subset of abnormal T cells involved in the pathogenesis of systemic autoimmunity in MRL-Faslpr and MRL-Fasgld mice. It plays a critical role in TCR and BCR signaling. The primary ligands for CD45 are galectin-1, CD2, CD3, and CD4. CD45R is commonly used as a pan-B cell marker; however, CD19 may be more appropriate for B cell specificity.

Product Details
Technical Data Sheet (pdf)

Product Details

Verified Reactivity
Mouse, Human
Reported Reactivity
Cat
Antibody Type
Monoclonal
Host Species
Rat
Immunogen
Abelson murine leukemia virus-induced pre-B tumor cells
Formulation
Phosphate-buffered solution, pH 7.2, containing 0.09% sodium azide and BSA (origin USA).
Preparation
The antibody was purified by affinity chromatography and conjugated with Brilliant Violet 510™ under optimal conditions.
Concentration
µg sizes: 0.2 mg/mL
µL sizes: lot-specific (to obtain lot-specific concentration and expiration, please enter the lot number in our Certificate of Analysis online tool.)
Storage & Handling
The antibody solution should be stored undiluted between 2°C and 8°C, and protected from prolonged exposure to light. Do not freeze.
Application

FC, IHC-F - Quality tested

SB - Reported in the literature, not verified in house
Recommended Usage

Each lot of this antibody is quality control tested by immunofluorescent staining with flow cytometric analysis. For flow cytometric staining using the µl size, the suggested use of this reagent is 5 µl per million cells in 100 µl staining volume or 5 µl per 100 µl of whole blood. For flow cytometric staining using the µg size, the suggested use of this reagent is ≤0.5 µg per million cells in 100 µl volume. It is recommended that the reagent be titrated for optimal performance for each application. It is recommended that the reagent be titrated for optimal performance for each application.

Brilliant Violet 510™ excites at 405 nm and emits at 510 nm. The bandpass filter 510/50 nm is recommended for detection, although filter optimization may be required depending on other fluorophores used. Be sure to verify that your cytometer configuration and software setup are appropriate for detecting this channel. Refer to your instrument manual or manufacturer for support. Brilliant Violet 510™ is a trademark of Sirigen Group Ltd.


Learn more about Brilliant Violet™.

This product is subject to proprietary rights of Sirigen Inc. and is made and sold under license from Sirigen Inc. The purchase of this product conveys to the buyer a non-transferable right to use the purchased product for research purposes only. This product may not be resold or incorporated in any manner into another product for resale. Any use for therapeutics or diagnostics is strictly prohibited. This product is covered by U.S. Patent(s), pending patent applications and foreign equivalents.
Excitation Laser
Violet Laser (405 nm)
Application Notes

Clone RA3-6B2 has been described to react with an epitope on the extracellular domain of the transmembrane CD45 glycoprotein which is dependent upon the expression of exon A and specific carbohydrate residues. Additional reported applications (for the relevant formats) include: immunoprecipitation1, in vitro and in vivo modulation of B cell responses2-4, immunohistochemistry of acetone-fixed frozen sections and formalin-fixed paraffin-embedded sections5,6, and spatial biology (IBEX)14,15.

Additional Product Notes

Iterative Bleaching Extended multi-pleXity (IBEX) is a fluorescent imaging technique capable of highly-multiplexed spatial analysis. The method relies on cyclical bleaching of panels of fluorescent antibodies in order to image and analyze many markers over multiple cycles of staining, imaging, and, bleaching. It is a community-developed open-access method developed by the Center for Advanced Tissue Imaging (CAT-I) in the National Institute of Allergy and Infectious Diseases (NIAID, NIH).

Application References
  1. Coffman RL. 1982. Immunol. Rev. 69:5. (IP)
  2. George A, et al. 1994. J. Immunol. 152:1014. (Activ)
  3. Asensi V, et al. 1989. Immunology 68:204. (Activ)
  4. Domiati-Saad R, et al. 1993. J. Immunol. 151:5936. (Activ)
  5. Hata H, et al. 2004. J. Clin. Invest. 114:582. (IHC)
  6. Monteith CE, et al. 1996. Can. J. Vet. Res. 60:193. (IHC)
  7. Shih FF, et al. 2006. J. Immunol. 176:3438. (FC)
  8. Chang C L-T, et al. 2007. J. Immunol. 178:6984.
  9. Fazilleau N, et al. 2007. Nature Immunol. 8:753.
  10. Lang GL, et al. 2008. Blood 111:2158. PubMed
  11. Charles N, et al. 2010. Nat. Med. 16:701. (FC) PubMed
  12. del Rio ML, et al. 2011. Transpl. Int. 24:501. (FC) PubMed
  13. Murakami R, et al. 2013. PLoS One. 8:73270. PubMed
  14. Radtke AJ, et al. 2020. Proc Natl Acad Sci U S A. 117:33455-65. (SB) PubMed
  15. Radtke AJ, et al. 2022. Nat Protoc. 17:378-401. (SB) PubMed
Product Citations
  1. Hutter K, et al. 2022. Front Immunol. 13:967914. PubMed
  2. Noir S, et al. 2017. Nucleic Acids Res. 10.1093/nar/gkx203. PubMed
  3. Tan J, et al. 2021. iScience. 24(8):102835. PubMed
  4. Vono M, et al. 2020. Cell Reports. 28(7):1773-1784.e5.. PubMed
  5. Shibata K, et al. 2022. Nat Commun. 13:6948. PubMed
  6. Lentucci C, et al. 2017. J Biol Chem. 292:2754-2772. PubMed
  7. Zhang J, et al. 2021. MedComm (Beijing). 2:256. PubMed
  8. Webb LMC, et al. 2021. Aging Cell. 20:e13295. PubMed
  9. Barry KC, et al. 2018. Nat Med. 24:1178. PubMed
  10. Shi B, et al. 2018. J Immunol. 200:586. PubMed
  11. Franks SE, et al. 2019. J Immunol. 202:3381. PubMed
  12. Xu AQ, et al. 2020. Elife. 9:00. PubMed
  13. Pan H, et al. 2020. Mol Psychiatry. . PubMed
  14. Yen WF et al. 2019. Cell reports. 27(5):1472-1486 . PubMed
  15. Huang L, et al. 2017. PLoS Biol.. 10.1371/journal.pbio.2001750. PubMed
  16. Yang L, et al. 2021. Cell Death Differ. 28:2616. PubMed
  17. Sato R, et al. 2020. Int Immunol. 499:32. PubMed
  18. Zbesko JC, et al. 2021. Brain Behav Immun. 91:578. PubMed
  19. Zhu Y, et al. 2022. Clin Transl Med. 12:e887. PubMed
  20. Hutter K, et al. 2020. FEBS J. . PubMed
  21. Webster P, et al. 2018. Nat Commun. 9:2649. PubMed
  22. Dietmar Herndler‐Brandstetter et al. 2018. Immunity. 48(4):716-729 . PubMed
  23. Mulas F, et al. 2020. Cell Mol Immunol. . PubMed
  24. Kurniawan H, et al. 2020. Cell Metabolism. 31(5):920-936. PubMed
  25. Schoeler K, et al. 2019. FEBS J. 10.1111/febs.14934. PubMed
  26. Reismann D, et al. 2017. Nat Commun.. 10.1038/s41467-017-01538-9. PubMed
  27. Chauveau L, et al. 2021. EMBO Rep. 22:e52447. PubMed
  28. Grioni M, et al. 2021. Blood Adv. 5:2817. PubMed
  29. Matundan H, et al. 2019. J Virol. 93. PubMed
  30. Walsh SM, et al. 2021. eLife. 10:00. PubMed
  31. Matsumura T, et al. 2022. Nat Commun. 13:7064. PubMed
  32. Lehrke MJ, et al. 2021. Elife. 10:. PubMed
  33. Zhang YN, et al. 2021. Sci Adv. 7:eabj3107. PubMed
  34. Schager AE, et al. 2018. MBio. 9:02379. PubMed
  35. Barbet G, et al. 2018. Immunity. 48:584. PubMed
  36. Molina MS, et al. 2021. Front Immunol. 12:699128. PubMed
  37. Yamada H, et al. 2018. J Immunol. 201:3492. PubMed
  38. Werner A, et al. 2021. iScience. 24:103076. PubMed
  39. Tähtinen S, et al. 2022. Nat Immunol. 23:532. PubMed
  40. Zhang Y, et al. 2022. Front Pharmacol. 13:952775. PubMed
  41. Rodrigues KA, et al. 2021. Sci Adv. 7:eabj6538. PubMed
  42. Orr MT, et al. 2019. NPJ Vaccines. 4:1. PubMed
  43. Jing Y, et al. 2019. J Allergy Clin Immunol. 144:1377. PubMed
  44. Bastow CR, et al. 2022. Front Immunol. 13:873586. PubMed
  45. Du Z, et al. 2020. J Allergy Clin Immunol. . PubMed
  46. Lucas ED, et al. 2020. Cell Reports. 33(2):108258. PubMed
  47. Campisi L, et al. 2016. Nat Immunol. 10.1038/ni.3512. PubMed
  48. Abdel Malik R, et al. 2017. Circ Res. 120:99. PubMed
RRID
AB_2561394 (BioLegend Cat. No. 103247)
AB_2650679 (BioLegend Cat. No. 103248)

Antigen Details

Structure
Protein tyrosine phosphatase (PTP) family, 180-240 kD
Distribution

B cells, T cell subset, NK cell subset

Function
Phosphatase, T and B cell activation
Ligand/Receptor
Galectin-1, CD2, CD3, CD4
Cell Type
B cells, NK cells, T cells
Biology Area
Cell Biology, Immunology, Inhibitory Molecules, Neuroscience, Neuroscience Cell Markers
Molecular Family
CD Molecules
Antigen References

1. Barclay A, et al. 1997. The Leukocyte Antigen FactsBook Academic Press.
2. Trowbridge IS, et al. 1993. Annu. Rev. Immunol. 12:85.
3. Kishihara K, et al. 1993. Cell 74:143.
4. Pulido R, et al. 1988. J. Immunol. 140:3851.

Gene ID
19264 View all products for this Gene ID 5788 View all products for this Gene ID
UniProt
View information about CD45R on UniProt.org

Related FAQs

If an antibody clone has been previously successfully used in IBEX in one fluorescent format, will other antibody formats work as well?

It’s likely that other fluorophore conjugates to the same antibody clone will also be compatible with IBEX using the same sample fixation procedure. Ultimately a directly conjugated antibody’s utility in fluorescent imaging and IBEX may be specific to the sample and microscope being used in the experiment. Some antibody clone conjugates may perform better than others due to performance differences in non-specific binding, fluorophore brightness, and other biochemical properties unique to that conjugate.

Will antibodies my lab is already using for fluorescent or chromogenic IHC work in IBEX?

Fundamentally, IBEX as a technique that works much in the same way as single antibody panels or single marker IF/IHC. If you’re already successfully using an antibody clone on a sample of interest, it is likely that clone will have utility in IBEX. It is expected some optimization and testing of different antibody fluorophore conjugates will be required to find a suitable format; however, legacy microscopy techniques like chromogenic IHC on fixed or frozen tissue is an excellent place to start looking for useful antibodies.

Are other fluorophores compatible with IBEX?

Over 18 fluorescent formats have been screened for use in IBEX, however, it is likely that other fluorophores are able to be rapidly bleached in IBEX. If a fluorophore format is already suitable for your imaging platform it can be tested for compatibility in IBEX.

The same antibody works in one tissue type but not another. What is happening?

Differences in tissue properties may impact both the ability of an antibody to bind its target specifically and impact the ability of a specific fluorophore conjugate to overcome the background fluorescent signal in a given tissue. Secondary stains, as well as testing multiple fluorescent conjugates of the same clone, may help to troubleshoot challenging targets or tissues. Using a reference control tissue may also give confidence in the specificity of your staining.

How can I be sure the staining I’m seeing in my tissue is real?

In general, best practices for validating an antibody in traditional chromogenic or fluorescent IHC are applicable to IBEX. Please reference the Nature Methods review on antibody based multiplexed imaging for resources on validating antibodies for IBEX.

Other Formats

View All CD45R/B220 Reagents Request Custom Conjugation
Description Clone Applications
Alexa Fluor® 594 anti-mouse/human CD45R/B220 RA3-6B2 IHC-F,FC,3D IHC
APC anti-mouse/human CD45R/B220 RA3-6B2 FC
Biotin anti-mouse/human CD45R/B220 RA3-6B2 FC,IHC-F
FITC anti-mouse/human CD45R/B220 RA3-6B2 FC
PE anti-mouse/human CD45R/B220 RA3-6B2 FC
PE/Cyanine5 anti-mouse/human CD45R/B220 RA3-6B2 FC
Purified anti-mouse/human CD45R/B220 RA3-6B2 FC,CyTOF®,IHC-F,IHC-P,IP,Activ
PE/Cyanine7 anti-mouse/human CD45R/B220 RA3-6B2 FC
APC/Cyanine7 anti-mouse/human CD45R/B220 RA3-6B2 FC
Alexa Fluor® 488 anti-mouse/human CD45R/B220 RA3-6B2 FC,IHC-F,3D IHC
Alexa Fluor® 647 anti-mouse/human CD45R/B220 RA3-6B2 FC,IHC-F,3D IHC,SB
Pacific Blue™ anti-mouse/human CD45R/B220 RA3-6B2 FC
Alexa Fluor® 700 anti-mouse/human CD45R/B220 RA3-6B2 FC
PerCP anti-mouse/human CD45R/B220 RA3-6B2 FC
PerCP/Cyanine5.5 anti-mouse/human CD45R/B220 RA3-6B2 FC
Brilliant Violet 421™ anti-mouse/human CD45R/B220 RA3-6B2 FC,IHC-F
Brilliant Violet 570™ anti-mouse/human CD45R/B220 RA3-6B2 FC
Brilliant Violet 650™ anti-mouse/human CD45R/B220 RA3-6B2 FC
Brilliant Violet 605™ anti-mouse/human CD45R/B220 RA3-6B2 FC
Brilliant Violet 785™ anti-mouse/human CD45R/B220 RA3-6B2 FC
Brilliant Violet 510™ anti-mouse/human CD45R/B220 RA3-6B2 FC,IHC-F,SB
Purified anti-mouse/human CD45R/B220 (Maxpar® Ready) RA3-6B2 FC,CyTOF®
Brilliant Violet 711™ anti-mouse/human CD45R/B220 RA3-6B2 FC
PE/Dazzle™ 594 anti-mouse/human CD45R/B220 RA3-6B2 FC
APC/Fire™ 750 anti-mouse/human CD45R/B220 RA3-6B2 FC
Brilliant Violet 750™ anti-mouse/human CD45R/B220 RA3-6B2 FC
TotalSeq™-A0103 anti-mouse/human CD45R/B220 RA3-6B2 PG
Spark Blue™ 550 anti-mouse/human CD45R/B220 RA3-6B2 FC
Spark NIR™ 685 anti-mouse/human CD45R/B220 RA3-6B2 FC
TotalSeq™-B0103 anti-mouse/human CD45R/B220 RA3-6B2 PG
Ultra-LEAF™ Purified anti-mouse/human CD45R/B220 RA3-6B2 FC,CyTOF®,IHC-F,IHC-P,IP,Activ
TotalSeq™-C0103 anti-mouse/human CD45R/B220 RA3-6B2 PG
PE/Fire™ 640 anti-mouse/human CD45R/B220 RA3-6B2 FC
APC/Fire™ 810 anti-mouse/human CD45R/B220 RA3-6B2 FC
PE/Fire™ 700 anti-mouse/human CD45R/B220 RA3-6B2 FC
Spark Violet™ 538 anti-mouse/human CD45R/B220 RA3-6B2 FC
Spark YG™ 581 anti-mouse/human CD45R/B220 RA3-6B2 FC
Spark YG™ 570 anti-mouse/human CD45R/B220 RA3-6B2 IHC-F
PE/Fire™ 810 anti-mouse/human CD45R/B220 RA3-6B2 FC
Spark Blue™ 574 anti-mouse/human CD45R/B220 Antibody RA3-6B2 FC
Spark Violet™ 423 anti-mouse/human CD45R/B220 Antibody RA3-6B2 FC
Spark Red™ 718 anti-mouse/human CD45R/B220 RA3-6B2 FC
Spark UV™ 387 anti-mouse/human CD45R/B220 RA3-6B2 FC
Spark PLUS UV™ 395 anti-mouse/human CD45R/B220 RA3-6B2 FC
Go To Top Version: 5    Revision Date: 04/22/2022

For Research Use Only. Not for diagnostic or therapeutic use.

 

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This data display is provided for general comparisons between formats.
Your actual data may vary due to variations in samples, target cells, instruments and their settings, staining conditions, and other factors.
If you need assistance with selecting the best format contact our expert technical support team.

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