Mouse B220/CD45R Antibody (RA3-6B2)

R&D Systems | Catalog # MAB1217

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Key Product Details

Species Reactivity

Validated:

Mouse

Cited:

Human, Mouse, Transgenic Mouse

Applications

Validated:

Flow Cytometry, Immunocytochemistry, Immunoprecipitation, CyTOF-reported

Cited:

Immunohistochemistry, Immunohistochemistry-Paraffin, Immunohistochemistry-Frozen, Flow Cytometry

Label

Unconjugated

Antibody Source

Monoclonal Rat IgG2A Clone # RA3-6B2
View all B220/CD45R Primary Antibodies »

Product Specifications

Immunogen

Abelson mouse leukemia virus-induced pre-B tumor cells

Specificity

Detects mouse B220/CD45R.

Clonality

Monoclonal

Host

Rat

Isotype

IgG2A

Scientific Data Images for Mouse B220/CD45R Antibody (RA3-6B2)

B220/CD45R antibody in Mouse Splenocytes by Immunocytochemistry (ICC).

B220/CD45R in Mouse Splenocytes.

B220/CD45R was detected in immersion fixed mouse splenocytes using Rat Anti-Mouse B220/CD45R Monoclonal Antibody (Catalog # MAB1217) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conju-gated Anti-Rat IgG Secondary Antibody (yellow; Catalog # NL013) and counter-stained with DAPI (blue). View our protocol for Fluorescent ICC Staining of Non-adherent Cells.
Detection of B220/CD45R antibody in Mouse Splenocytes antibody by Flow Cytometry.

Detection of B220/CD45R in Mouse Splenocytes by Flow Cytometry.

Mouse splenocytes were stained with Rat Anti-Mouse CD3 APC-conjugated Monoclonal Antibody (Catalog # FAB4841A) and either (A) Rat Anti-Mouse B220/CD45R Monoclonal Antibody (Catalog # MAB1217) or (B) Rat IgG2AIsotype Control (Catalog # MAB006) followed by Phycoerythrin-conjugated Anti-Rat IgG Secondary Antibody (Catalog # F0105B).
Detection of B220/CD45R by Flow Cytometry

Detection of B220/CD45R by Flow Cytometry

pDCs downregulate Siglec-H upon MCMV infection in a TLR9- and MyD88-dependent manner in vitro.Flt3-L derived total BMDCs or CD11c+B220+ pDCs sorted from BMDC cultures were mock treated with PBS, CpG treated or MCMV infected at MOI 2 unless indicated otherwise. (A) Representative FACS plots show Siglec-H expression and MCMV-GFP expression of BMDCs (upper panel). Representative histogram plots display B220 expression on CD11c+ Siglec-H+ DCs at 0, 3, 6 and 24 h p.i. compared with mock treated controls (lower panel). (B) Representative histogram plots display Siglec-H expression of sorted wt, TLR9−/− and MyD88−/− pDCs after stimulation or infection. (C) Representative histogram plots display Siglec-H expression of sorted wt and IFNAR−/− pDCs 24 h after infection or stimulation. (D) Quantification of the Siglec-H expression from wt and IFNAR−/− pDCs after MCMV infection relative to their respective mock controls. Data are from one of three individual experiments with similar results. *p<0.05, ***p<0.001, Students t-test. (Gray bar = isotype control, gray dashed line = mock treatment and dark bold line = CpG-A treatment or MCMV infection). Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/24086137), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of B220/CD45R by Flow Cytometry

Detection of B220/CD45R by Flow Cytometry

pDCs downregulate Siglec-H upon MCMV infection in a TLR9- and MyD88-dependent manner in vitro.Flt3-L derived total BMDCs or CD11c+B220+ pDCs sorted from BMDC cultures were mock treated with PBS, CpG treated or MCMV infected at MOI 2 unless indicated otherwise. (A) Representative FACS plots show Siglec-H expression and MCMV-GFP expression of BMDCs (upper panel). Representative histogram plots display B220 expression on CD11c+ Siglec-H+ DCs at 0, 3, 6 and 24 h p.i. compared with mock treated controls (lower panel). (B) Representative histogram plots display Siglec-H expression of sorted wt, TLR9−/− and MyD88−/− pDCs after stimulation or infection. (C) Representative histogram plots display Siglec-H expression of sorted wt and IFNAR−/− pDCs 24 h after infection or stimulation. (D) Quantification of the Siglec-H expression from wt and IFNAR−/− pDCs after MCMV infection relative to their respective mock controls. Data are from one of three individual experiments with similar results. *p<0.05, ***p<0.001, Students t-test. (Gray bar = isotype control, gray dashed line = mock treatment and dark bold line = CpG-A treatment or MCMV infection). Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/24086137), licensed under a CC-BY license. Not internally tested by R&D Systems.

Applications for Mouse B220/CD45R Antibody (RA3-6B2)

Application
Recommended Usage

CyTOF-reported

Lee, H. et al. (2015) Mucosal Immunol. 8: 1083. Ready to be labeled using established conjugation methods. No BSA or other carrier proteins that could interfere with conjugation.

Flow Cytometry

0.25 µg/106 cells
Sample: Mouse splenocytes

Immunocytochemistry

8-25 µg/mL
Sample: Immersion fixed mouse splenocytes

Immunoprecipitation

Coffman, R.L. (1982) Immuno. Rev. 69:5.

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Flow Cytometry

Formulation, Preparation, and Storage

Purification

Protein A or G purified from hybridoma culture supernatant

Reconstitution

Reconstitute at 0.5 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

Stability & Storage

Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 6 months, -20 to -70 °C under sterile conditions after reconstitution.

Calculators

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Background: B220/CD45R

B220 is the mouse CD45 isoform CD45R, which is predominantly expressed on all B lymphocytes, including pro-, mature and activated B cells (1‑3). The level of B220 antigen expression on the B cell lineage is developmentally regulated (2‑4) and this antibody is commonly used as a B cell marker. RA3-6B2 has been reported to inhibit in vivo B cell responses (5, 6).

References

  1. Coffman, R.L. (1982) Immuno. Rev. 69:5.
  2. Hardy, R.R. et al. (1991) J. Exp. Med. 173:1213.
  3. Hathcock, K.S. et al. (1992) J. Immunol. 149:2286.
  4. Allman, D.M. et al. (1992) J. Immunol. 149:2533.
  5. Asensi, V.K. et al. (1989) Immunology 68:204.
  6. Domiati-Saad, R. et al. (1993) J. Immunol. 151:5936.

Long Name

Cluster of Differentiation 45

Alternate Names

CD45R

Entrez Gene IDs

5788 (Human); 19264 (Mouse); 24699 (Rat)

Gene Symbol

PTPRC

Additional B220/CD45R Products

Product Documents for Mouse B220/CD45R Antibody (RA3-6B2)

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Product Specific Notices for Mouse B220/CD45R Antibody (RA3-6B2)

For research use only

Citations for Mouse B220/CD45R Antibody (RA3-6B2)

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Protocols

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