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Multicolor flow cytometric analysis of CD20 expression on mouse splenocytes. Mouse splenocytes from C57BL/6 mice were preincubated with Mouse BD Fc Block™ (Cat. No. 553141/553142). 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 CD20 antibody (Cat No. 567250; Right Plot) at 0.5 ug/test. The bivariate pseudocolor density plots showing correlated expression of CD20 (or Ig Isotype control staining) versus CD45R/B220 signal were derived from gated events with the forward and side light-scatter characteristics of viable mouse splenocytes. Flow cytometric analysis was performed using a BD FACSCelesta™ Flow Cytometer System and FlowJo software™. Data shown on this Technical Data Sheet are not lot specific.
BD Pharmingen™ PE Rat Anti-Mouse CD20
Regulatory Status Legend
Any use of products other than the permitted use without the express written authorization of Becton, Dickinson and Company is strictly prohibited.
Preparation And Storage
Recommended Assay Procedures
BD™ CompBeads can be used as surrogates to assess fluorescence spillover (Compensation). When fluorochrome conjugated antibodies are bound to 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 cell and CompBead to ensure that BD Comp beads are appropriate for your specific cellular application.
Product Notices
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- An isotype control should be used at the same concentration as the antibody of interest.
- 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.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
The GOT214A monoclonal antibody specifically binds to mouse CD20, also known as B-lymphocyte antigen and Bp35. CD20, a member of the MS4A gene family, is a 33-37 kDa protein comprised of four transmembrane spanning regions that present as a homo-oligomeric complexes on the cell surface. CD20 is expressed on most mature B cells but not on plasma cells. During development, CD20 is initially expressed after the induction of CD19 and is co-expressed with IgM during the pre-B to immature B cell transition in bone marrow. Expression of CD20 increases during maturation with almost all mature B cells expressing CD20 at some level. Unlike other B cell antigens, CD20 is not shed or internalized upon antibody binding. CD20 regulates B-cell activation and proliferation. Its ligation promotes transmembrane Ca2+ trafficking.
Development References (2)
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Uchida J, Hamaguchi Y, Oliver JA, et al. The innate mononuclear phagocyte network depletes B lymphocytes through Fc receptor-dependent mechanisms during anti-CD20 antibody immunotherapy.. J Exp Med. 2004; 199(12):1659-1669. (Biology). View Reference
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Uchida J, Lee Y, Hasegawa M, et al. Mouse CD20 expression and function.. Int Immunol. 2004; 16(1):119-29. (Biology). View Reference
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For Research Use Only. Not for use in diagnostic or therapeutic procedures.