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PE Mouse Anti-Human HLA-A3
PE Mouse Anti-Human HLA-A3
Multiparameter flow cytometric analysis of HLA-A3 expression on human peripheral blood leucocyte populations. Human whole blood was stained with either PE Mouse IgG2a, κ Isotype Control (Cat. No. 554648; Left Plot) or PE Mouse Anti-Human HLA-A3 antibody (Cat. No. 566605; Right Plot) at 0.5 µg/test. Erythrocytes were lysed with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899). Two-parameter flow cytometric contour plots showing the correlated expression of HLA-A3 (or Ig Isotype control staining) versus side light-scatter (SSC-A) signals were derived from gated events with the forward and side light-scatter characteristics of viable leucocyte populations. Flow cytometric analysis was performed using a BD LSRFortessa™ X-20 Flow Cytometer System.
PE Mouse Anti-Human HLA-A3

Flow cytometric analysis of HLA-A3 expression on Jurkat cells. Jurkat cells were stained with PE Mouse IgG2a, κ Isotype Control (dashed line histogram) or PE Mouse Anti-Human HLA-A3 antibody (solid line histogram) at 0.5 µg/test. The fluorescence histogram showing HLA-A3 expression (or Ig Isotype control staining) was derived from gated events with the forward and side light-scatter characteristics of viable cells. Flow cytometric analysis was performed using a BD LSRFortessa™ X-20 Flow Cytometer System. Data shown on this Technical Data Sheet are not lot specific.

Multiparameter flow cytometric analysis of HLA-A3 expression on human peripheral blood leucocyte populations. Human whole blood was stained with either PE Mouse IgG2a, κ Isotype Control (Cat. No. 554648; Left Plot) or PE Mouse Anti-Human HLA-A3 antibody (Cat. No. 566605; Right Plot) at 0.5 µg/test. Erythrocytes were lysed with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899). Two-parameter flow cytometric contour plots showing the correlated expression of HLA-A3 (or Ig Isotype control staining) versus side light-scatter (SSC-A) signals were derived from gated events with the forward and side light-scatter characteristics of viable leucocyte populations. Flow cytometric analysis was performed using a BD LSRFortessa™ X-20 Flow Cytometer System.

Flow cytometric analysis of HLA-A3 expression on Jurkat cells. Jurkat cells were stained with PE Mouse IgG2a, κ Isotype Control (dashed line histogram) or PE Mouse Anti-Human HLA-A3 antibody (solid line histogram) at 0.5 µg/test. The fluorescence histogram showing HLA-A3 expression (or Ig Isotype control staining) was derived from gated events with the forward and side light-scatter characteristics of viable cells. Flow cytometric analysis was performed using a BD LSRFortessa™ X-20 Flow Cytometer System. Data shown on this Technical Data Sheet are not lot specific.

Product Details
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BD Pharmingen™
HLA class I histocompatibility antigen, A-3 alpha chain; HLA Antigen A3
Human (QC Testing)
Mouse BALB/c IgG2a, κ
Human Lymphocytes from Mixed Lymphocyte Reaction
Flow cytometry (Routinely Tested)
0.2 mg/ml
AB_2739760
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 with R-PE under optimum conditions, and unconjugated antibody and free PE were removed.

Product Notices

  1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
  2. An isotype control should be used at the same concentration as the antibody of interest.
  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. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  5. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
566605 Rev. 1
Antibody Details
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GAP.A3

The GAP.A3 monoclonal antibody specifically recognizes the polymorphic HLA class I histocompatibility antigen, A-3 alpha chain (HLA-A3). This ~44 kDa human major histocompatibility complex (MHC) class I molecule noncovalently associates with the ~12 kDa monomorphic β2 microglobulin. Two major subtypes are encoded by HLA-A3, HLA-A3.1 and HLA-A3.2. HLA-A3 gene expression is more commonly detected in individuals from Europe and southern India. HLA-A3 is expressed on nearly all nucleated cells of HLA-A3-positive individuals. HLA-A3 functions in the presentation of antigens to CD8-positive T cells which may lead to the generation of HLA-A3-restricted cytotoxic T cells and memory T cells. When complexed with certain antigens, HLA-A3 can also bind to killer cell immunoglobulin-like receptors (KIR), such as KIR3DL2, and might regulate NK cell function.

Note: Since HLA-A3 expression varies between human populations, clone GAP.A3 staining can be donor-dependent. Based on in-house testing and current literature, individuals of European or Southern Indian descent more frequently express HLA-A3 than those of Asian descent. Data may differ between donors due to geographical variations of HLA-A3 expression.

566605 Rev. 1
Format Details
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PE
R-Phycoerythrin (PE), is part of the BD family of Phycobiliprotein dyes. This fluorochrome is a multimeric fluorescent phycobiliprotein with excitation maximum (Ex Max) of 496 nm and 566 nm and an emission maximum (Em Max) at 576 nm. PE is designed to be excited by the Blue (488 nm), Green (532 nm) and Yellow-Green (561 nm) lasers and detected using an optical filter centered near 575 nm (e.g., a 575/26-nm bandpass filter). As PE is excited by multiple lasers, this can result in cross-laser excitation and fluorescence spillover on instruments with various combinations of Blue, Green, and Yellow-Green lasers. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
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PE
Yellow-Green 488 nm, 532 nm, 561 nm
496 nm, 566 nm
576 nm
566605 Rev.1
Citations & References
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Development References (7)

  1. Allen TM, Altfeld M, Yu XG, et al. Selection, transmission, and reversion of an antigen-processing cytotoxic T-lymphocyte escape mutation in human immunodeficiency virus type 1 infection.. J Virol. 2004; 78(13):7069-78. (Biology). View Reference
  2. Augusto DG, O'Connor GM, Lobo-Alves SC, et al. Pemphigus is associated with KIR3DL2 expression levels and provides evidence that KIR3DL2 may bind HLA-A3 and A11 in vivo.. Eur J Immunol. 2015; 45(7):2052-60. (Biology). View Reference
  3. Berger AE, Davis JE, Cresswell P. Monoclonal antibody to HLA-A3.. Hybridoma. 1982; 1(2):87-90. (Immunogen: Cytotoxicity, Immunoprecipitation). View Reference
  4. Carr TM, Adair SJ, Fink MJ, Hogan KT. Immunological profiling of a panel of human ovarian cancer cell lines.. Cancer Immunol Immunother. 2008; 57(1):31-42. (Clone-specific: Flow cytometry). View Reference
  5. Hansasuta P, Dong T, Thananchai H, et al. Recognition of HLA-A3 and HLA-A11 by KIR3DL2 is peptide-specific.. Eur J Immunol. 2004; 34(6):1673-9. (Biology). View Reference
  6. Lalouel JM, Jorde LB. Idiopathic hemochromatosis: significance and implications of linkage and association to HLA.. Ann N Y Acad Sci. 1988; 526:34-46. (Biology). View Reference
  7. Vonderheide RH, Anderson KS, Hahn WC, Butler MO, Schultze JL, Nadler LM. Characterization of HLA-A3-restricted cytotoxic T lymphocytes reactive against the widely expressed tumor antigen telomerase.. Clin Cancer Res. 2001; 7(11):3343-8. (Biology). View Reference
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566605 Rev. 1

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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.