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PE Rat Anti-Mouse CD21/CD35
PE Rat Anti-Mouse CD21/CD35
Flow cytometric analysis of CD21/CD35 expression on mouse splenic leucocytes. Mouse splenocytes from C57BL/6 mice were preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™) (Cat. No. 553141/553142). The cells were stained with APC Rat Anti-Mouse CD45R/B220 antibody (Cat No. 553092/561880) and either PE Rat IgG2a, κ Isotype Control (Cat No. 553930, Left Plot) or PE Rat Anti-Mouse CD21/CD35 antibody (Cat No. 568014; Right Plot) at 0.25 µg/test. The bivariate pseudocolor density plot showing the correlated expression of CD21/CD35 (or Ig Isotype control staining) versus CD45R/B220 was derived from gated events with the forward and side light-scatter characteristics of viable mouse splenocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa™ Cell Analyzer System and FlowJo™ software. Data shown on this Technical Data Sheet are not lot specific.
Flow cytometric analysis of CD21/CD35 expression on mouse splenic leucocytes. Mouse splenocytes from C57BL/6 mice were preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™) (Cat. No. 553141/553142). The cells were stained with APC Rat Anti-Mouse CD45R/B220 antibody (Cat No. 553092/561880) and either PE Rat IgG2a, κ Isotype Control (Cat No. 553930, Left Plot) or PE Rat Anti-Mouse CD21/CD35 antibody (Cat No. 568014; Right Plot) at 0.25 µg/test. The bivariate pseudocolor density plot showing the correlated expression of CD21/CD35 (or Ig Isotype control staining) versus CD45R/B220 was derived from gated events with the forward and side light-scatter characteristics of viable mouse splenocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa™ Cell Analyzer System and FlowJo™ software. Data shown on this Technical Data Sheet are not lot specific.
Product Details
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BD Pharmingen™
CD21/CD35; CR2/CR1; mCR1/mCR2
Mouse (QC Testing)
Rat SD, also known as Sprague-Dawley (outbred) IgG2a, κ
Purified Mouse CR1 Protein
Flow cytometry (Routinely Tested)
0.2 mg/ml
Aqueous buffered solution containing protein stabilizer and ≤0.09% sodium azide.
RUO


Preparation And Storage

Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze. The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. The antibody was conjugated to the dye under optimum conditions and unconjugated antibody and free dye were removed.

Recommended Assay Procedures

BD® CompBeads can be used as surrogates to assess fluorescence spillover (Compensation). When fluorochrome conjugated antibodies are bound to BD® CompBeads, they have spectral properties very similar to cells. However, for some fluorochromes there can be small differences in spectral emissions compared to cells, resulting in spillover values that differ when compared to biological controls. It is strongly recommended that when using a reagent for the first time, users compare the spillover on cells and BD CompBeads to ensure that BD® CompBeads are appropriate for your specific cellular application.

Product Notices

  1. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  2. Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
  3. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  4. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
  5. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
  6. An isotype control should be used at the same concentration as the antibody of interest.
568014 Rev. 1
Antibody Details
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7E9

The 7E9 antibody specifically recognizes CD21/CD35, an epitope shared by mouse Complement receptor 2 (CR2) and Complement receptor 1 (CR1), which are also known as CD21 and CD35, respectively. Mouse CD21 and CD35 are ~145-kDa and ~190-kDa single pass type I transmembrane glycoproteins, respectively. Unlike human CD21 and CD35 that are encoded by separate CR2 and CR1 genes, mouse CD21 and CD35 are encoded by a single Cr2 gene and are generated by alternative mRNA splicing. CD21 and CD35 are expressed on B cells and follicular dendritic cells (FCD) but not on thymocytes, T cells, erythrocytes, or platelets. CD35 is also expressed on macrophages and activated granulocytes. CD21 binds to cleaved C3b fragments, including C3d- and C3dg-bound to antigen, whereas CD35 binds to C3b-, iC3b-, and C4b-bound complexes. These receptors play many important roles including the transport, presentation, and retention of complement fragment-tagged antigens and immune complexes crucial for generating strong B cell antibody-producing responses. Complement-tagged antigen can bind to CD21, part of the B cell coreceptor complex and to the B cell receptor for antigen (BCR) which facilitates the antigen-driven activation of B cell responses. CD35 interacts with other regulatory factors to promote C3b and C4b degradation as well as playing roles in stimulating phagocytic functions in activated leucocytes. The 7E9 antibody recognizes an epitope on CD35 distinct from the epitope recognized by the mouse CD35-specific clone, 8C12, as well as the mouse CD21/CD35-specific clone, 7G6.

568014 Rev. 1
Format Details
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PE
R-Phycoerythrin (PE), is part of the BD family of Phycobiliprotein dyes. This fluorochrome is a multimeric fluorescent phycobiliprotein with excitation maximum (Ex Max) of 496 nm and 566 nm and an emission maximum (Em Max) at 576 nm. PE is designed to be excited by the Blue (488 nm), Green (532 nm) and Yellow-Green (561 nm) lasers and detected using an optical filter centered near 575 nm (e.g., a 575/26-nm bandpass filter). As PE is excited by multiple lasers, this can result in cross-laser excitation and fluorescence spillover on instruments with various combinations of Blue, Green, and Yellow-Green lasers. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
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PE
Yellow-Green 488 nm, 532 nm, 561 nm
496 nm, 566 nm
576 nm
568014 Rev.1
Citations & References
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Development References (6)

  1. Ahearn JM, Fischer MB, Croix D, et al. Disruption of the Cr2 locus results in a reduction in B-1a cells and in an impaired B cell response to T-dependent antigen. Immunity. 1996; 4(3):251-262. (Clone-specific: Flow cytometry). View Reference
  2. Heyman B, Wiersma EJ, Kinoshita T. In vivo inhibition of the antibody response by a complement receptor-specific monoclonal antibody. J Exp Med. 1990; 172(2):665-668. (Clone-specific: In vivo exacerbation). View Reference
  3. Jacobson AC, Weis JH. Comparative functional evolution of human and mouse CR1 and CR2. J Immunol. 2008; 181(5):2953-2959. (Biology). View Reference
  4. Kinoshita T, Takeda J, Hong K, Kozono H, Sakai H, Inoue K. Monoclonal antibodies to mouse complement receptor type 1 (CR1). Their use in a distribution study showing that mouse erythrocytes and platelets are CR1-negative. J Immunol. 1988; 140(9):3066-3072. (Immunogen: Flow cytometry, Immunoprecipitation, Radioimmunoassay). View Reference
  5. Kinoshita T, Thyphronitis G, Tsokos GC, et al. Characterization of murine complement receptor type 2 and its immunological cross-reactivity with type 1 receptor. Int Immunol. 1990; 2(7):651-659. (Clone-specific: Flow cytometry). View Reference
  6. Molina H, Holers VM, Li B, et al. Markedly impaired humoral immune response in mice deficient in complement receptors 1 and 2. Proc Natl Acad Sci U S A. 1996; 93(8):3357-3361. (Clone-specific: Flow cytometry). View Reference
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568014 Rev. 1

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