Alexa Fluor® 594 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
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Product Citations
publications
1_RA3-6B2_AF594_B220_Antibody_IHC_042314
C57BL/6 mouse frozen lymph node section was 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 2.5 µg/mL of B220 (clone RA3-6B2) Alexa Fluor® 594 (red), 2.5 µg/mL of CD4 (clone RM4-5) Alexa Fluor® 488 (green), and 2.5 µg/mL of CD8 (clone 53-6.7) Alexa Fluor® 647 (blue) overnight at 4°C. The image was captured by 10X objective.
  • 1_RA3-6B2_AF594_B220_Antibody_IHC_042314
    C57BL/6 mouse frozen lymph node section was 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 2.5 µg/mL of B220 (clone RA3-6B2) Alexa Fluor® 594 (red), 2.5 µg/mL of CD4 (clone RM4-5) Alexa Fluor® 488 (green), and 2.5 µg/mL of CD8 (clone 53-6.7) Alexa Fluor® 647 (blue) overnight at 4°C. The image was captured by 10X objective.
  • 2_RA3-6B2_AF594_CD45R_Antibody_FC_072414.jpg
    C57BL/6 mouse splenocytes were stained with CD45R/B220 (clone RA3-6B2) Alexa Fluor® 594 (filled histogram) or rat IgG2a, κ Alexa Fluor® 594 isotype control (open histogram). The data was acquired by BD LSRFortessa™ cell analyzer equipped with Yellow-Green Laser (561 nm).
  • 3_Still-image_0401-5_10X-1_1
    Formalin-fixed, 300 micron-thick mouse spleen section was blocked, permeabilized and stained overnight with CD4 (clone RM4-5) Alexa Fluor® 647 (red), CD169 (Siglec-1)(clone 3D6.112) Alexa Fluor® 488 (green), and CD45R/B220 (clone RA3-6B2) Alexa Fluor® 594 (blue) all at 5 µg/mL, optically cleared, then analyzed at 220 μm imaging depth on a confocal microscope. Watch the video.
  • 4_500A2_A594_CD3epsilon_Antibody_3_091721.png
    Paraformaldehyde-fixed (4%), 500 μm-thick mouse spleen section was processed according to the Ce3DTM Tissue Clearing Kit protocol (Cat. No. 427701). The section was costained with anti-mouse CD3ε Antibody (clone 500A2) Alexa Fluor® 488 at 5 µg/mL (green), and anti-mouse/human CD45R/B220 Antibody (clone RA3-6B2) Alexa Fluor® 594 at 5 µg/mL (magenta) and counterstained with DAPI (blue). The section was then optically cleared and mounted in a sample chamber. The image was captured with a 10X objective using Zeiss 780 confocal microscope and processed by Imaris image analysis software.
    Watch the video.
Compare all formats See Alexa Fluor® 594 spectral data See high resolution IHC data...
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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

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® 594 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

IHC-F - Quality tested
FC, 3D IHC - Verified

Recommended Usage

Each lot of this antibody is quality control tested by immunofluorescence staining. For immunohistochemistry, a concentration range of 2.5-5 μg/mL is recommended. For immunofluorescence microscopy, a concentration range of 1.25 - 10 μg/mL is recommended. For flow cytometric staining, the suggested use of this reagent is ≤0.25 µg per million cells in 100 µL volume. For 3D immunohistochemistry on formalin-fixed tissues, a concentration of 5.0 µg/mL is suggested. It is recommended that the reagent be titrated for optimal performance for each application.

* Alexa Fluor® 594 has an excitation maximum of 590 nm, and a maximum emission of 617 nm.


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

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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.

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. Han B, et al. 2022. Hepatology. 76:357. PubMed
  2. Koenig A, et al. 2022. Front Immunol. 12:791100. PubMed
  3. Ariyasu Y, et al. 2022. Int J Mol Sci. 23: . PubMed
  4. Baptista AP et al. 2019. Immunity. 50(5):1188-1201 . PubMed
  5. Li S, et al. 2020. Cell Death Dis. 0.9875. PubMed
  6. Pham THM, et al. 2020. Cell Host & Microbe. 27(1):54-67.e5.. PubMed
  7. Counoupas C, et al. 2020. NPJ Vaccines. 0.28125. PubMed
  8. Matryba P, et al. 2020. J Immunol. 1395:204. PubMed
  9. Wang X, et al. 2021. EMBO J. 40:e105926. PubMed
  10. Kobia FM, et al. 2020. PLoS Biol. 18:e3000850. PubMed
  11. Luo L, et al. 2021. J Neuroinflammation. 18:27. PubMed
  12. Shi H et al. 2018. Immunity. 49(5):899-914 . PubMed
  13. Song W, et al. 2022. Immunity. 55:290. PubMed
  14. Goldberg MF et al. 2018. Immunity. 49(6):1090-1102 . PubMed
  15. Kuhn NF et al. 2019. Cancer cell. 35(3):473-488 . PubMed
  16. Bidet K, et al. 2019. NPJ Vaccines. 4:27. PubMed
RRID
AB_2563229 (BioLegend Cat. No. 103254)

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

There are no FAQs for this product.

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 UV395™ anti-mouse/human CD45R/B220 RA3-6B2 FC
Go To Top Version: 5    Revision Date: 09.17.2021

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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