C57BL/6 mouse slenocytes were stained with purified B220 (clone RA3-6B2) (filled histogram) or rat IgG2a, κ (open histogram), followed by anti-rat IgG FITC.
C57BL/6 mouse slenocytes were stained with purified B220 (clone RA3-6B2) (filled histogram) or rat IgG2a, κ (open histogram), followed by anti-rat IgG FITC.
C57BL/6 mouse frozen spleen section was fixed with 4% paraformaldehyde (PFA) for ten minutes at room temperature and blocked with 5% FBS for 30 minutes at room temperature. Then, the section was stained with 10 µg/mL of purified anti-mouse/human CD45R/B220 (clone RA3-6B2) and anti-mouse CD3ε (clone 500A2) Alexa Fluor® 594 (red) overnight at 4°C, followed by 2.5 µg/mL of anti-rat IgG (clone Poly4054) Alexa Fluor® 488 (green) for two hours at room temperature. Nuclei were counterstained with DAPI (blue). The image was captured by 10X objective.
Fresh, frozen mouse spleen was stained with purified CD45R/B220 clone Ra3-6B2 conjugated and detected with a FITC CODEX™ oligonucleotide duplex (red). Samples were counterstained with TCT A647 (green). Data generated at Akoya Biosciences, Inc. using the CODEX™ technology.
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.
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 106 cells in 100 µl volume. For immunohistochemistry on frozen tissue sections, a concentration range of 5.0 - 10 µg/ml is suggested. It is recommended that the reagent be titrated for optimal performance for each application.
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
Coffman RL. 1982. Immunol. Rev. 69:5. (IP)
George A, et al. 1994. J. Immunol. 152:1014. (Activ)
Asensi V, et al. 1989. Immunology 68:204. (Activ)
Domiati-Saad R, et al. 1993. J. Immunol. 151:5936. (Activ)
Hata H, et al. 2004. J. Clin. Invest. 114:582. (IHC)
Monteith CE, et al. 1996. Can. J. Vet. Res. 60:193. (IHC)
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.
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.
C57BL/6 mouse slenocytes were stained with purified B220 (clone RA3-6B2) (filled histogram) or rat IgG2a, κ (open histogram), followed by anti-rat IgG FITC.
C57BL/6 mouse slenocytes were stained with purified B220 (clone RA3-6B2) (filled histogram) or rat IgG2a, κ (open histogram), followed by anti-rat IgG FITC.
C57BL/6 mouse frozen spleen section was fixed with 4% paraformaldehyde (PFA) for ten minutes at room temperature and blocked with 5% FBS for 30 minutes at room temperature. Then, the section was stained with 10 µg/mL of purified anti-mouse/human CD45R/B220 (clone RA3-6B2) and anti-mouse CD3ε (clone 500A2) Alexa Fluor® 594 (red) overnight at 4°C, followed by 2.5 µg/mL of anti-rat IgG (clone Poly4054) Alexa Fluor® 488 (green) for two hours at room temperature. Nuclei were counterstained with DAPI (blue). The image was captured by 10X objective.
Fresh, frozen mouse spleen was stained with purified CD45R/B220 clone Ra3-6B2 conjugated and detected with a FITC CODEX™ oligonucleotide duplex (red). Samples were counterstained with TCT A647 (green). Data generated at Akoya Biosciences, Inc. using the CODEX™ technology.
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.
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 106 cells in 100 µl volume. For immunohistochemistry on frozen tissue sections, a concentration range of 5.0 - 10 µg/ml is suggested. It is recommended that the reagent be titrated for optimal performance for each application.
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
Coffman RL. 1982. Immunol. Rev. 69:5. (IP)
George A, et al. 1994. J. Immunol. 152:1014. (Activ)
Asensi V, et al. 1989. Immunology 68:204. (Activ)
Domiati-Saad R, et al. 1993. J. Immunol. 151:5936. (Activ)
Hata H, et al. 2004. J. Clin. Invest. 114:582. (IHC)
Monteith CE, et al. 1996. Can. J. Vet. Res. 60:193. (IHC)
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.
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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