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Flow cytometric analysis of HLA-A2 expression on Human lymphocytes from HLA-A2-positive and -negative donors. Human whole blood from either an HLA-A2-negative (Left Plot) or an HLA-A2-positive (Right Plot) donor was stained with BD Horizon™ BV421 Mouse IgG1, κ Isotype Control (Cat No. 562438: dashed line histograms) or BD Horizon™ BV421 Mouse Anti-Human HLA-A2 antibody (Cat. No. 568757; solid line histograms) at 0.5 µg/test. Erythrocytes were lysed with BD FACS Lysing™ Solution (Cat. No. 349202). The fluorescence histograms showing HLA-A2 expression (or Ig Isotype control staining) were 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. Data shown on this Technical Data Sheet are not lot specific.
BD Horizon™ BV421 Mouse Anti-Human HLA-A2
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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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Companion Products
The monoclonal antibody BB7.2 specifically binds to the α subunit of the human leukocyte antigen-A2 (HLA-A2), a class I molecule of the major histocompatibility complex (MHC). The MHC gene locus encodes a group of highly polymorphic, cell-surface proteins that play a broad role in the immune response to protein antigens. MHC molecules bind and present small antigenic protein fragments to antigen-specific receptors expressed by T cells (TCR). Human (human leukocyte antigen/HLA) MHC molecules are comprised of two major classes, MHC class I and class II. Functionally, class I MHC molecules bind peptides derived from intracellular antigens (eg, viral and some bacterial antigens) which are specifically recognized by CD8+ T cells. Class II MHC molecules bind antigens derived from pathogens multiplying in intracellular vesicles and ingested extracellular bacteria, both of which are recognized by CD4+ T cells. TCR recognize processed peptides bound to the MHC as well as regions of the MHC molecule itself. CD4 and CD8 accessory molecules strengthen the formation of the TCR-MHC complex through their interaction with non-polymorphic regions of the MHC molecule.
Development References (4)
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Bjorkman PJ, Saper MA, Samraoui B, Bennett WS, Strominger JL, Wiley DC. Structure of the human class I histocompatibility antigen, HLA-A2. Nature. 1987; 329(6139):506-512. (Biology). View Reference
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Bjorkman PJ, Saper MA, Samraoui B, Bennett WS, Strominger JL, Wiley DC. The foreign antigen binding site and T cell recognition regions of class I histocompatibility antigens. Nature. 1987; 329(6139):512-518. (Biology). View Reference
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Parham P, Brodsky FM. Partial purification and some properties of BB7.2. A cytotoxic monoclonal antibody with specificity for HLA-A2 and a variant of HLA-A28. Hum Immunol. 1981; 3(4):277-299. (Clone-specific: Blocking, Cytotoxicity, Inhibition). View Reference
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Romero P, Dunbar PR, Valmori D. Ex vivo staining of metastatic lymph nodes by class I major histocompatibility complex tetramers reveals high numbers of antigen-experienced tumor-specific cytolytic T lymphocytes. J Exp Med. 1998; 188(9):1641-1650. (Biology). View Reference
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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.
Refer to manufacturer's instructions for use and related User Manuals and Technical Data Sheets before using this product 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.