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Alexa Fluor™ 488 Rat Anti-Mouse CD20
Alexa Fluor™ 488 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™ BUV395 Rat Anti-Mouse CD45R/B220 antibody (Cat. No. 563793) and with either Alexa Fluor™ 488 Mouse IgG2a, κ Isotype Control (Cat No. 557676; Left Plot) or Alexa Fluor™ 488 Rat Anti-Mouse CD20 antibody (Cat No. 568246/568247; Right Plot) at 1.0 ug/test. DAPI (4',6-Diamidino-2-Phenylindole, Dihydrochloride) Solution (Cat. No. 564907) 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 (DAPI-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™ BUV395 Rat Anti-Mouse CD45R/B220 antibody (Cat. No. 563793) and with either Alexa Fluor™ 488 Mouse IgG2a, κ Isotype Control (Cat No. 557676; Left Plot) or Alexa Fluor™ 488 Rat Anti-Mouse CD20 antibody (Cat No. 568246/568247; Right Plot) at 1.0 ug/test. DAPI (4',6-Diamidino-2-Phenylindole, Dihydrochloride) Solution (Cat. No. 564907) 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 (DAPI-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 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 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. Alexa Fluor® 488 fluorochrome emission is collected at the same instrument settings as for fluorescein isothiocyanate (FITC).
  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. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
  5. An isotype control should be used at the same concentration as the antibody of interest.
  6. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  7. This product is provided under an intellectual property license between Life Technologies Corporation and BD Businesses. The purchase of this product conveys to the buyer the non-transferable right to use the purchased amount of the product and components of the product in research conducted by the buyer (whether the buyer is an academic or for-profit entity). The buyer cannot sell or otherwise transfer (a) this product (b) its components or (c) materials made using this product or its components to a third party or otherwise use this product or its components or materials made using this product or its components for Commercial Purposes. Commercial Purposes means any activity by a party for consideration and may include, but is not limited to: (1) use of the product or its components in manufacturing; (2) use of the product or its components to provide a service, information, or data; (3) use of the product or its components for therapeutic, diagnostic or prophylactic purposes; or (4) resale of the product or its components, whether or not such product or its components are resold for use in research. For information on purchasing a license to this product for any other use, contact Life Technologies Corporation, Cell Analysis Business Unit Business Development, 29851 Willow Creek Road, Eugene, OR 97402, USA, Tel: (541) 465-8300. Fax: (541) 335-0504.
  8. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
  9. Alexa Fluor™ is a trademark of Life Technologies Corporation.
568247 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.

568247 Rev. 1
Format Details
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Alexa Fluor™ 488
Alexa Fluor™ 488 Dye is part of the BD blue family of dyes. This is a small organic fluorochrome with an excitation maximum (Ex Max) at 494-nm and an emission maximum (Em Max) at 517-nm. Alexa Fluor™ 488 is designed to be excited by the Blue laser (488 nm) and detected using an optical filter centered near 520-nm (e.g., a 530/30-nm bandpass filter). Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
Alexa Fluor™ 488
Blue 488 nm
494 nm
517 nm
568247 Rev.1
Citations & References
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View product citations for antibody "568247" 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
568247 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.