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BV421 Mouse Anti-Human CD20
BV421 Mouse Anti-Human CD20

Multiparameter flow cytometric analysis of CD20 expression on human peripheral blood leucocyte populations. Whole blood was stained with either BD Horizon™ BV421 Mouse IgG2b, κ Isotype Control (Cat. No. 562748; Left Plot) or with BD Horizon™ BV421 Mouse Anti-Human CD20 antibody (Cat. No. 562873; Right Plot). Erythrocytes were lysed with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899). Bivariate pseudocolor density plots showing the correlated expression of CD20 (or Ig Isotype Control staining) versus side light-scatter (SSC-A) derived from gated events with the forward light-scatter characteristics of intact leucocytes. Flow cytometry and data analysis was performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ software.

Multiparameter flow cytometric analysis of CD20 expression on human peripheral blood leucocyte populations. Whole blood was stained with either BD Horizon™ BV421 Mouse IgG2b, κ Isotype Control (Cat. No. 562748; Left Plot) or with BD Horizon™ BV421 Mouse Anti-Human CD20 antibody (Cat. No. 562873; Right Plot). Erythrocytes were lysed with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899). Bivariate pseudocolor density plots showing the correlated expression of CD20 (or Ig Isotype Control staining) versus side light-scatter (SSC-A) derived from gated events with the forward light-scatter characteristics of intact leucocytes. Flow cytometry and data analysis was performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ software.

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_2737857
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 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 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.

For optimal and reproducible results, BD Horizon Brilliant™ Stain Buffer should be used anytime BD Horizon Brilliant™ dyes are used in a multicolor flow cytometry panel. Fluorescent dye interactions may cause staining artifacts which may affect data interpretation. The BD Horizon Brilliant™ Stain Buffer was designed to minimize these interactions. When BD Horizon Brilliant™ Stain Buffer is used in in the multicolor panel, it should also be used in the corresponding compensation controls for all dyes to achieve the most accurate compensation. For the most accurate compensation, compensation controls created with either cells or beads should be exposed to BD Horizon Brilliant™ Stain Buffer for the same length of time as the corresponding multicolor panel. More information can be found in the Technical Data Sheet of the BD Horizon Brilliant™ Stain Buffer (Cat. No. 563794/566349) or the BD Horizon Brilliant™ Stain Buffer Plus (Cat. No. 566385).

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. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  6. BD Horizon Brilliant Violet 421 is covered by one or more of the following US patents: 8,158,444; 8,362,193; 8,575,303; 8,354,239.
  7. BD Horizon Brilliant Stain Buffer is covered by one or more of the following US patents: 8,110,673; 8,158,444; 8,575,303; 8,354,239.
  8. Species cross-reactivity detected in product development may not have been confirmed on every format and/or application.
  9. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  10. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
  11. Pacific Blue™ is a trademark of Life Technologies Corporation.
562873 Rev. 3
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.

This clone also cross-reacts with a subset of peripheral blood lymphocytes, but not monocytes nor granulocytes, of baboon and both rhesus and cynomolgus macaque monkeys. The distribution on lymphocytes is similar to that seen with normal human donor lymphocytes, namely bright staining on B lymphocytes and weak reactivity on a small subset of CD3-positive T lymphocytes.

The antibody was conjugated to BD Horizon™ BV421 which is part of the BD Horizon Brilliant™ Violet family of dyes. With an Ex Max near 407 nm and Em Max near 421 nm, BD Horizon™ BV421 can be excited by the violet laser (405 nm) and detected with a 450/50 nm filter. BD Horizon™ BV421 conjugates are very bright, often exhibiting a 10 fold improvement in brightness compared to Pacific Blue™ conjugates. Due to nearly identical excitation and emission properties but different spillover characteristics, BD Horizon™ BV421, Pacific Blue™, and BD Horizon™ V450 cannot be used simultaneously.

562873 Rev. 3
Format Details
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BV421
The BD Horizon Brilliant Violet™ 421 (BV421) Dye is part of the BD Horizon Brilliant Violet™ family of dyes. This polymer-technology based dye has an excitation maximum (Ex Max) of 407-nm and an emission maximum (Em Max) at 423-nm. Driven by BD innovation, BV421 is designed to be excited by the violet laser (405-nm) and detected using an optical filter centered near 420-nm (e.g., a 431/28-nm or 450/50-nm bandpass filter). BV421 is an ideal alternative for V450 as it is approximately ten times brighter with less spillover into the BV510/V500 detector. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
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BV421
Violet 405 nm
407 nm
423 nm
562873 Rev.3
Citations & References
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Development References (7)

  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. 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
  6. Reinherz EL. Ellis L. Reinherz .. et al., ed. Leukocyte typing II. New York: Springer-Verlag; 1986:1-560.
  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.
View All (7) View Less
562873 Rev. 3

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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.