Alexa Fluor® 647 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_AF647_090507
C57BL/6 mouse splenocytes stained with RA3-6B2 Alexa Fluor® 647
  • 1_RA3-6B2_AF647_090507
    C57BL/6 mouse splenocytes stained with RA3-6B2 Alexa Fluor® 647
  • 2_RA3-6B2_A647_B220_Antibody_IHC-F_080519
    C57BL/6 mouse frozen lymph node sections were fixed with 4% paraformaldehyde (PFA) for 10 minutes at room temperature and blocked with 5% FBS plus 5% rat serum for 1 hour at room temperature. Then the section was stained with 5 µg/mL of CD4 (clone GK1.5) Alexa Fluor® 594 (red), 5 µg/mL of B220 (clone RA3-6B2) Alexa Fluor® 647 (green) overnight at 4°C. The image was captured by 10X objective.
  • 3_Still-image_0316-5_20X-1_1
    Formalin-fixed, 300 micron-thick mouse spleen section was blocked, permeabilized and stained overnight with CD3 (clone 17A2) Alexa Fluor® 488 (red), CD21/35 (CR2/CR1)(clone 7E9) Alexa Fluor® 594 (green), and CD45R/B220 (clone RA3-6B2) Alexa Fluor® 647 (blue) all at 5 µg/mL, optically cleared, then analyzed at 215 μm imaging depth on a confocal microscope.
Compare all formats See Alexa Fluor® 647 spectral data
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103229 25 µg 76€
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103226 100 µg 172€
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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.
Preparation
The antibody was purified by affinity chromatography and conjugated with Alexa Fluor® 647 under optimal conditions.
Concentration
0.5 mg/mL
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 - Quality tested
IHC-F, 3D IHC - Verified

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, the suggested use of this reagent is ≤ 0.25 µg per million cells in 100 µL volume. For immunohistochemistry on frozen tissue sections, a concentration range of 2.5 - 5.0 µg/mL is suggested. For immunofluorescence microscopy, a concentration range of 1.25 - 10 µg/mL is recommended. It is recommended that the reagent be titrated for optimal performance for each application.

* Alexa Fluor® 647 has a maximum emission of 668 nm when it is excited at 633nm / 635nm.


Alexa Fluor® and Pacific Blue™ are trademarks of Life Technologies Corporation.

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Excitation Laser
Red Laser (633 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

(PubMed link indicates BioLegend citation)
  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. Gil-Cruz C, et al. 2012. Proc Natl Acad Sci U S A. 109:1233. PubMed
  2. Harumiya S, et al. 2013. Arch Biochem Biophys. 533:18. PubMed
  3. Folgosa L, et al. 2013. J Immunol. 191:5951. PubMed
  4. Medgyesi D, et al. 2014. J Exp Med. 211:427. PubMed
  5. Neubert K, et al. 2014. J Immunol. 192:5830. PubMed
  6. Hoechst B, et al. 2016. J Immunol. 195:1517-1523. PubMed
  7. Kang S, et al. 2016. J Immunol. 196: 196 - 206. PubMed
  8. Yoshikawa S, et al. 2016. Sci Rep. 6:18738. PubMed
  9. Campisi L, et al. 2016. Nat Immunol. 10.1038/ni.3512. PubMed
  10. Prados A, Kollias G, Koliaraki V 2016. Sci Rep. 6: 33027. PubMed
  11. Hu HJ, et al. 2020. Cell Death Dis. 1.168055556. PubMed
  12. Cabañero D, et al. 2020. Elife. 9:00. PubMed
  13. Kim EH, et al. 2020. Elife. 9:00. PubMed
  14. Dammeijer F, et al. 2020. Cancer Cell. 38(5):685-700.e8. PubMed
  15. Emgård J, et al. 2018. Immunity. 143:419. PubMed
  16. Lin JR et al. 2018. eLife. 7 pii: e31657. PubMed
  17. Beura LK, et al. 2018. Immunity. 48:327. PubMed
  18. Hermetet F, et al. 2019. Nat Commun. 10:523. PubMed
  19. Ramakrishnan SK, et al. 2019. Nat Commun. 10:660. PubMed
  20. Cheng HW, et al. 2019. Nat Commun. 10:1739. PubMed
  21. Barry KC, et al. 2018. Nat Med. 24:1178. PubMed
  22. Farsakoglu Y et al. 2019. Cell reports. 26(9):2307-2315 . PubMed
  23. Simula L et al. 2018. Cell reports. 25(11):3059-3073 . PubMed
  24. Liu D et al. 2019. Immunity. 51(1):64-76 . PubMed
  25. Magri G et al. 2017. Immunity. 47(1):118-134 . PubMed
  26. Tsai F, et al. 2017. J Exp Med. 214:3753. PubMed
  27. Fallet B, et al. 2020. Cell Rep. 30:1013. PubMed
  28. LaFleur MW, et al. 2019. Nat Immunol. 20:1335. PubMed
  29. Sarapulov AV, et al. 2020. Front Immunol. 11:599. PubMed
  30. Chou YJ, et al. 2020. Sci Rep. 10:8422. PubMed
  31. Chauveau A, et al. 2020. Immunity. 52:794. PubMed
  32. Cao A, et al. 2018. Nat Commun. 9:3288. PubMed
  33. Yang J, et al. 2019. FASEB J. 33:12780. PubMed
  34. Lian J, et al. 2020. Cell Reports. 31(8):107679. PubMed
  35. Brownlie R, et al. 2008. J Exp Med. 205:883. PubMed
RRID
AB_492875 (BioLegend Cat. No. 103229)
AB_389330 (BioLegend Cat. No. 103226)

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
Go To Top Version: 7    Revision Date: 04.22.2022

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*These products may be covered by one or more Limited Use Label Licenses (see the BioLegend Catalog or our website, www.biolegend.com/ordering#license). BioLegend products may not be transferred to third parties, resold, modified for resale, or used to manufacture commercial products, reverse engineer functionally similar materials, or to provide a service to third parties without written approval of BioLegend. By use of these products you accept the terms and conditions of all applicable Limited Use Label Licenses. Unless otherwise indicated, these products are for research use only and are not intended for human or animal diagnostic, therapeutic or commercial 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.

  • Alexa Fluor® 594 anti-mouse/human CD45R/B220

  • APC anti-mouse/human CD45R/B220

  • Biotin anti-mouse/human CD45R/B220

  • FITC anti-mouse/human CD45R/B220

  • PE anti-mouse/human CD45R/B220

  • PE/Cyanine5 anti-mouse/human CD45R/B220

  • Purified anti-mouse/human CD45R/B220

  • PE/Cyanine7 anti-mouse/human CD45R/B220

  • APC/Cyanine7 anti-mouse/human CD45R/B220

  • Alexa Fluor® 488 anti-mouse/human CD45R/B220

  • Alexa Fluor® 647 anti-mouse/human CD45R/B220

  • Pacific Blue™ anti-mouse/human CD45R/B220

  • Alexa Fluor® 700 anti-mouse/human CD45R/B220

  • PerCP anti-mouse/human CD45R/B220

  • PerCP/Cyanine5.5 anti-mouse/human CD45R/B220

  • Brilliant Violet 421™ anti-mouse/human CD45R/B220

  • Brilliant Violet 570™ anti-mouse/human CD45R/B220

  • Brilliant Violet 650™ anti-mouse/human CD45R/B220

  • Brilliant Violet 605™ anti-mouse/human CD45R/B220

  • Brilliant Violet 785™ anti-mouse/human CD45R/B220

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

  • Purified anti-mouse/human CD45R/B220 (Maxpar® Ready)

  • Brilliant Violet 711™ anti-mouse/human CD45R/B220

  • PE/Dazzle™ 594 anti-mouse/human CD45R/B220

  • APC/Fire™ 750 anti-mouse/human CD45R/B220

  • Brilliant Violet 750™ anti-mouse/human CD45R/B220

  • TotalSeq™-A0103 anti-mouse/human CD45R/B220

  • Spark Blue™ 550 anti-mouse/human CD45R/B220

  • Spark NIR™ 685 anti-mouse/human CD45R/B220

  • TotalSeq™-B0103 anti-mouse/human CD45R/B220

  • Ultra-LEAF™ Purified anti-mouse/human CD45R/B220

  • TotalSeq™-C0103 anti-mouse/human CD45R/B220

  • PE/Fire™ 640 anti-mouse/human CD45R/B220

  • APC/Fire™ 810 anti-mouse/human CD45R/B220

  • PE/Fire™ 700 anti-mouse/human CD45R/B220

  • Spark Violet™ 538 anti-mouse/human CD45R/B220

  • Spark YG™ 581 anti-mouse/human CD45R/B220

  • Spark YG™ 570 anti-mouse/human CD45R/B220

  • PE/Fire™ 810 anti-mouse/human CD45R/B220

  • Spark Blue™ 574 anti-mouse/human CD45R/B220 Antibody

  • Spark Violet™ 423 anti-mouse/human CD45R/B220 Antibody

  • Spark Red™ 718 anti-mouse/human CD45R/B220

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