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R718 Rat Anti-Mouse CD20
R718 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 BD Horizon™ BV421 Rat Anti-Mouse CD45R/B220 antibody (Cat. No. 562922) and with either BD Horizon™ R718 Rat IgG2a, κ Isotype Control (Cat No. 566941; Left Plot) or BD Horizon™ R718 Rat Anti-Mouse CD20 antibody (Cat No. 568258/568259; Right Plot) at 1.0 ug/test. BD Via-Probe™ Cell Viability 7-AAD Solution (Cat. No. 555815/555816) was added to cells right before analysis. The bivariate pseudocolor density plot showing the correlated expression of CD20 (or Ig Isotype control staining) versus CD45R/B220 signal was derived from gated events with the forward and side light-scatter characteristics of viable (7-AAD-negative) mouse splenocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer 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 BD Horizon™ BV421 Rat Anti-Mouse CD45R/B220 antibody (Cat. No. 562922) and with either BD Horizon™ R718 Rat IgG2a, κ Isotype Control (Cat No. 566941; Left Plot) or BD Horizon™ R718 Rat Anti-Mouse CD20 antibody (Cat No. 568258/568259; Right Plot) at 1.0 ug/test. BD Via-Probe™ Cell Viability 7-AAD Solution (Cat. No. 555815/555816) was added to cells right before analysis. The bivariate pseudocolor density plot showing the correlated expression of CD20 (or Ig Isotype control staining) versus CD45R/B220 signal was derived from gated events with the forward and side light-scatter characteristics of viable (7-AAD-negative) mouse splenocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ software. Data shown on this Technical Data Sheet are not lot specific.
Product Details
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BD Horizon™
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 unreacted dye was 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. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
  4. An isotype control should be used at the same concentration as the antibody of interest.
  5. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
  6. This product is provided under an Agreement between BIOTIUM and BD Biosciences. This product, and only in the amount purchased by buyer, may be used solely for buyer’s own internal research, in a manner consistent with the accompanying product literature. No other right to use, sell or otherwise transfer (a) this product, or (b) its components is hereby granted expressly, by implication or by estoppel. This product is for research use only. Diagnostic uses require a separate license from Biotium, Inc. For information on purchasing a license to this product including for purposes other than research, contact Biotium, Inc., 3159 Corporate Place, Hayward, CA 94545, Tel: (510) 265-1027. Fax: (510) 265-1352. Email: btinfo@biotium.com.
  7. Alexa Fluor™ is a trademark of Life Technologies Corporation.
568259 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.

568259 Rev. 1
Format Details
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R718
The BD Horizon™ Red 718 (R718) Dye is part of the BD red family of dyes. It is a small organic fluorochrome with an excitation maximum (Ex Max) at 695-nm and an emission maximum (Em Max) at 718-nm. Driven by BD innovation, R718 is designed to be excited by the red laser (627–640-nm) and detected using an optical filter centered near 720-nm (e.g., a 720/40-nm bandpass filter). R718 is a brighter alternative to Alexa Fluor™ 700. R718 is also a bright small molecule alternative to APC-R700 with lower spread into the APC detector. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
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R718
Red 627-640 nm
695 nm
718 nm
568259 Rev.1
Citations & References
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View product citations for antibody "568259" on CiteAb

Development References (3)

  1. Tedder TF, Engel P. CD20: a regulator of cell-cycle progression of B lymphocytes.. Immunol Today. 1994; 15(9):450-4. (Biology). View Reference
  2. 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: Flow cytometry). View Reference
  3. Uchida J, Lee Y, Hasegawa M, et al. Mouse CD20 expression and function.. Int Immunol. 2004; 16(1):119-29. (Biology). View Reference
568259 Rev. 1

 

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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.