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BV480 Mouse Anti-Human CD20
BV480 Mouse Anti-Human CD20
Two-color flow cytometric analysis of CD20 expression on human peripheral blood lymphocytes. Whole blood was stained with APC Mouse Anti-Human CD19 antibody (Cat. No. 555415/561742) and either BD Horizon™ BV480 Mouse IgG2b, κ Isotype Control (Cat. No. 566080; Left Plot) or BD Horizon BV480 Mouse Anti-Human CD20 antibody (Cat. No. 566132/566181; Right Plot). Erythrocytes were lysed with BD FACS Lysing Solution (Cat. No. 349202).  The two-color flow cytometric contour plot showing the correlated expression of CD20 (or Ig Isotype control staining) versus CD19 was derived from gated events with the forward and side light-scatter characteristics of intact lymphocytes. Flow cytometric analysis was performed using a BD LSRFortessa™ Cell Analyzer System.
Two-color flow cytometric analysis of CD20 expression on human peripheral blood lymphocytes. Whole blood was stained with APC Mouse Anti-Human CD19 antibody (Cat. No. 555415/561742) and either BD Horizon™ BV480 Mouse IgG2b, κ Isotype Control (Cat. No. 566080; Left Plot) or BD Horizon BV480 Mouse Anti-Human CD20 antibody (Cat. No. 566132/566181; Right Plot). Erythrocytes were lysed with BD FACS Lysing Solution (Cat. No. 349202).  The two-color flow cytometric contour plot showing the correlated expression of CD20 (or Ig Isotype control staining) versus CD19 was derived from gated events with the forward and side light-scatter characteristics of intact lymphocytes. Flow cytometric analysis was performed using a BD LSRFortessa™ Cell Analyzer System.
Product Details
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BD Horizon™
MS4A1; B1; Bp35; LEU-16; S7
Human (QC Testing), Rhesus, Cynomolgus, Baboon (Tested in Development)
Mouse C57BL/6 IgG2b, κ
Human 6.16c1.3 B cell line
Flow cytometry (Routinely Tested)
5 µl
II B22; III B739, NL382; IV B201
931
AB_2739531
Aqueous buffered solution containing BSA 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 with BD Horizon BV480 under optimum conditions, and unconjugated antibody and free BD Horizon BV480 were removed.

Recommended Assay Procedures

For optimal and reproducible results, BD Horizon Brilliant Stain Buffer should be used anytime two or more BD Horizon Brilliant dyes are used in the same experiment. Fluorescent dye interactions may cause staining artifacts which may affect data interpretation. The BD Horizon Brilliant Stain Buffer was designed to minimize these interactions. More information can be found in the Technical Data Sheet of the BD Horizon Brilliant Stain Buffer (Cat. No. 563794).

Product Notices

  1. This reagent has been pre-diluted for use at the recommended Volume per Test. We typically use 1 × 10^6 cells in a 100-µl experimental sample (a test).
  2. An isotype control should be used at the same concentration as the antibody of interest.
  3. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
  4. 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.
  5. Species cross-reactivity detected in product development may not have been confirmed on every format and/or application.
  6. BD Horizon Brilliant Violet 480 is covered by one or more of the following US patents: 8,575,303; 8,354,239.
  7. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  8. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
566181 Rev. 1
Antibody Details
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2H7

The 2H7 monoclonal antibody specifically binds to CD20, encoded by the MS4A1 (Membrane-spanning 4-domains, subfamily A, member 1) gene. CD20 is a 33-37 kDa, unglycosylated four-transmembrane phosphoprotein. CD20 is expressed on pre-B-cells, resting and activated B cells, and follicular dendritic cells, but not plasma cells. Low level CD20 expression is observed on a small subset of normal circulating T lymphocytes. The CD20 molecule is involved in the regulation of B-cell activation.

The antibody was conjugated to BD Horizon BV480 which is part of the BD Horizon Brilliant™ Violet family of dyes. With an Ex Max of 436-nm and Em Max at 478-nm, BD Horizon BV480 can be excited by the violet laser and detected in the BD Horizon BV510 (525/40-nm) filter set.  BV480 has less spillover into the BV605 detector and, in general, is brighter than BV510.

566181 Rev. 1
Format Details
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BV480
The BD Horizon Brilliant Violet™ 480 (BV480) Dye is part of the BD Horizon Brilliant Violet™ family of dyes. This polymer-technology fluorochrome has an excitation maximum (Ex Max) of 440-nm and an emission maximum (Em Max) of 479-nm. Driven by BD innovation, BV480 is designed to be excited by the violet laser (405-nm) and detected using an optical filter centered near 480-nm (e.g., a 525/50 bandpass filter). The increased fluorescence intensity of BV480 and narrower emission spectra, make it a good alternative for BV510 or V500. Due to its excitation profile, BV480 will also has less cross-laser excitation with the UV laser, resulting in less spillover into UV channels compared to BV510. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
altImg
BV480
Violet 405 nm
440 nm
479 nm
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Citations & References
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Development References (8)

  1. Clark EA, Yokochi T. Human B cell and B cell blast-associated surface molecules defined with monoclonal antibodies. In: Bernard A, Boumsell L, Dausset J, Milstein C, Schlossman SF, ed. Leukocyte Typing. Berlin: Springer-Verlag; 1984:339-346.
  2. Hultin LE, Hausner MA, Hultin PM, Giorgi JV. CD20 (pan-B cell) antigen is expressed at a low level on a subpopulation of human T lymphocytes. Cytometry. 1993; 14(2):193-204. (Biology). View Reference
  3. Knapp W. W. Knapp .. et al., ed. Leucocyte typing IV : white cell differentiation antigens. Oxford New York: Oxford University Press; 1989:1-1182.
  4. Ledbetter JA, Clark EA. Surface phenotype and function of tonsillar germinal center and mantle zone B cell subsets. Hum Immunol. 1986; 15:30-43. (Immunogen: Blocking, Flow cytometry). View Reference
  5. Lin G-X, Yang X, Hollemweguer E, et al. Cross-reactivity of CD antibodies in eight animal species. In: Mason D. David Mason .. et al., ed. Leucocyte typing VII : white cell differentiation antigens : proceedings of the Seventh International Workshop and Conference held in Harrogate, United Kingdom. Oxford: Oxford University Press; 2002:519-523.
  6. Loken MR, Shah VO, Dattilio KL, Civin CI. Flow cytometric analysis of human bone marrow. II. Normal B lymphocyte development. Blood. 1987; 70(5):1316-1324. (Biology). View Reference
  7. Schlossman SF. Stuart F. Schlossman .. et al., ed. Leucocyte typing V : white cell differentiation antigens : proceedings of the fifth international workshop and conference held in Boston, USA, 3-7 November, 1993. Oxford: Oxford University Press; 1995.
  8. Tedder TF, Penta A. Structure of the CD20 antigen and gene of human and mouse B cells: use of transfected cell lines to examine the Workshop panel of antibodies. In: Knapp W. W. Knapp .. et al., ed. Leucocyte typing IV : white cell differentiation antigens. Oxford New York: Oxford University Press; 1989:48-50.
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For Research Use Only. Not for use in diagnostic or therapeutic procedures.