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Multiparameter flow cytometric analysis of CD79a expression in Human peripheral mononuclear cells. Human peripheral mononuclear cells (PBMC) were fixed with BD Cytofix™ Fixation Buffer (Cat. No. 554655) and permeabilized with BD Perm/Wash™ Buffer (Cat. No. 554723). The cells were subsequently stained with either BD Horizon™ BV421 Mouse IgG1, κ Isotype Control (Cat. No. 569394; Left Plot) or BD Horizon™ BV421 Mouse Anti-Human CD79a antibody (Cat. No. 570950/571021; Right Plot). The bivariate pseudocolor density plot showing the correlated expression of CD79a (or Ig Isotype control staining) versus side light-scatter (SSC-A) signals was derived from gated events with the forward and side light-scatter characteristics of intact PBMC. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ Software.
Flow cytometric analysis of CD79a expression in Human peripheral blood lymphocytes. Human peripheral blood mononuclear cells (PBMC) were fixed with BD Cytofix™ Fixation Buffer (Cat. No. 554655) and permeabilized with BD Perm/Wash™ Buffer (Cat. No. 554723). The cells were subsequently stained with either BD Horizon™ BV421 Mouse IgG1, κ Isotype Control (Cat. No. 569394; dashed line histogram) or BD Horizon™ BV421 Mouse Anti-Human CD79a antibody (Cat. No. 570950/571021; solid line histogram). The fluorescence histogram showing CD79a expression (or Ig Isotype control staining) was derived from gated events with the forward and side light-scatter characteristics of intact lymphocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ Software.
BD Horizon™ BV421 Mouse Anti-Human CD79a
BD Horizon™ BV421 Mouse Anti-Human CD79a
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Preparation And Storage
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).
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- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- 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).
- An isotype control should be used at the same concentration as the antibody of interest.
- 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.
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Companion Products
The HM57 monoclonal antibody specifically binds to the cytoplasmic domain of CD79a. CD79a is a 47-kDa type 1 transmembrane glycoprotein present on B lymphocytes. CD79a is also referred to as mb-1, IGA and Ig-alpha (Ig-α). It is expressed on B cells at various stages of differentiation, from the pre-B cell stage, probably before expression of cytoplasmic μ chain, to the plasma cell stage, in which it is detected only in the cytoplasm. CD79a associates with CD79b to form part of the B-cell receptor complex. It has been suggested that CD79a may play a role in mediating the transport of IgM to the cell surface. This antibody reportedly may crossreact with monkey, mouse, rat, bovine, canine, chicken, equine, guinea pig, porcine, and rabbit CD79a.
Development References (6)
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Engel P, Wagner N, Tedder TF. CD79 Workshop report. In: 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; 1995:667-670.
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Faldyna M, Sinkora J, Knotigova P, Rehakova Z, Moravkova A, Toman M. Flow cytometric analysis of bone marrow leukocytes in neonatal dogs. Vet Immunol Immunopathol. 2003; 95(3-4):165-176. (Clone-specific: Flow cytometry). View Reference
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Jones M, Cordell JL, Beyers AD, Tse AG, Mason DY. Detection of T and B cells in many animal species using cross-reactive anti-peptide antibodies. J Immunol. 1993; 150(12):5429-5435. (Clone-specific: Immunohistochemistry, Western blot). View Reference
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Mason DY, Cordell JL, Tse AG, et al. The IgM-associated protein mb-1 as a marker of normal and neoplastic B cells. J Immunol. 1991; 147(11):2474-2482. (Immunogen: Flow cytometry, Immunoaffinity chromatography, Immunofluorescence, Immunohistochemistry, Immunoprecipitation). View Reference
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Sakaguchi N, Kashiwamura S, Kimoto M, Thalmann P, Melchers F. B lymphocyte lineage-restricted expression of mb-1, a gene with CD3-like structural properties. EMBO J. 1988; 7(11):3457-3464. (Biology). View Reference
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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.
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For Research Use Only. Not for use in diagnostic or therapeutic procedures.