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PE Rat Anti-Mouse CD20
PE Rat Anti-Mouse CD20

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.

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.

Product Details
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BD Pharmingen™
Ly-44; Ms4a1; Ms4a2
Mouse (QC Testing)
Rat IgG2a, κ
RBL-1-mCD20 transfected cells
Flow cytometry (Routinely Tested)
0.2 mg/ml
Aqueous buffered solution containing ≤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 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

  1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
  2. An isotype control should be used at the same concentration as the antibody of interest.
  3. 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.
  4. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  5. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
  6. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
567250 Rev. 1
Antibody Details
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GOT214A

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.

567250 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
567250 Rev.1
Citations & References
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Development References (2)

  1. 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
  2. Uchida J, Lee Y, Hasegawa M, et al. Mouse CD20 expression and function.. Int Immunol. 2004; 16(1):119-29. (Biology). View Reference
567250 Rev. 1

Please refer to Support Documents for Quality Certificates


Global - Refer to manufacturer's instructions for use and related User Manuals and Technical data sheets before using this products as described


Comparisons, where applicable, are made against older BD Technology, manual methods or are general performance claims.  Comparisons are not made against non-BD technologies, unless otherwise noted.

For Research Use Only. Not for use in diagnostic or therapeutic procedures.

Refer to manufacturer's instructions for use and related User Manuals and Technical Data Sheets before using this product as described.

Comparisons, where applicable, are made against older BD technology, manual methods or are general performance claims. Comparisons are not made against non-BD technologies, unless otherwise noted.