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Purified Mouse Anti-Human HLA-ABC
Purified Mouse Anti-Human HLA-ABC

Analyses of HLA-ABC Expression

       Left Panel - Multiparameter flow cytometric analysis of human HLA-ABC expression on peripheral blood leucocytes. Whole blood cells were lysed and fixed with BD Phosflow™ Lyse/Fix Buffer (Cat. No. 558049), washed, and then stained with either Purified Mouse IgG1 κ Isotype Control (Cat. No. 554121; Left Plot) or Purified Mouse Anti-Human HLA-ABC antibody (Cat. No. 565292; Right Plot). The cells were further stained with PE Goat Anti-Mouse Ig (Multiple Adsorption) (Cat. No. 550589). Flow cytometric contour plots showing the correlated expression of HLA-ABC (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 intact leucocyte populations. Flow cytometric analysis was performed using a BD LSRFortessa™ system.

       Right Panel - Immunohistochemical staining of HLA-ABC expression in human renal carcinoma tissue. Following antigen retrieval with BD Retrievagen A Buffer (Cat. No. 550524), the formalin-fixed paraffin-embedded sections were stained with either Purified Mouse IgG1 κ Isotype Control (Left Image) or Purified Mouse HLA-ABC antibody (Right Image). A three-step staining procedure that employs a Biotin Goat Anti-Mouse Immunoglobulin (Cat. No. 550337), Streptavidin-Horseradish Peroxidase (HRP) (Cat. No.550946), and the DAB Substrate Kit (Cat. No. 550880) was used to develop the primary staining reagents. As shown in the Right Image, the human HLA-ABC-specific antibody primarily surface stained some renal carcinoma cells. Original magnification: 40×.

Analyses of HLA-ABC Expression

       Left Panel - Multiparameter flow cytometric analysis of human HLA-ABC expression on peripheral blood leucocytes. Whole blood cells were lysed and fixed with BD Phosflow™ Lyse/Fix Buffer (Cat. No. 558049), washed, and then stained with either Purified Mouse IgG1 κ Isotype Control (Cat. No. 554121; Left Plot) or Purified Mouse Anti-Human HLA-ABC antibody (Cat. No. 565292; Right Plot). The cells were further stained with PE Goat Anti-Mouse Ig (Multiple Adsorption) (Cat. No. 550589). Flow cytometric contour plots showing the correlated expression of HLA-ABC (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 intact leucocyte populations. Flow cytometric analysis was performed using a BD LSRFortessa™ system.

       Right Panel - Immunohistochemical staining of HLA-ABC expression in human renal carcinoma tissue. Following antigen retrieval with BD Retrievagen A Buffer (Cat. No. 550524), the formalin-fixed paraffin-embedded sections were stained with either Purified Mouse IgG1 κ Isotype Control (Left Image) or Purified Mouse HLA-ABC antibody (Right Image). A three-step staining procedure that employs a Biotin Goat Anti-Mouse Immunoglobulin (Cat. No. 550337), Streptavidin-Horseradish Peroxidase (HRP) (Cat. No.550946), and the DAB Substrate Kit (Cat. No. 550880) was used to develop the primary staining reagents. As shown in the Right Image, the human HLA-ABC-specific antibody primarily surface stained some renal carcinoma cells. Original magnification: 40×.

Product Details
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BD Pharmingen™
HLA-A, HLA-B, HLA-C; HLAA, HLAB, HLAC; HLA-A,B,C
Human (QC Testing)
Mouse BALB/c IgG1, κ
Human HLA-A Recombinant Protein
Flow cytometry (Routinely Tested), Immunohistochemistry (Tested During Development)
0.5 mg/ml
AB_2739161
Aqueous buffered solution containing ≤0.09% sodium azide.
RUO


Preparation And Storage

Store undiluted at 4°C. The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography.

Product Notices

  1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
  2. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  3. Sodium azide is a reversible inhibitor of oxidative metabolism; therefore, antibody preparations containing this preservative agent must not be used in cell cultures nor injected into animals. Sodium azide may be removed by washing stained cells or plate-bound antibody or dialyzing soluble antibody in sodium azide-free buffer. Since endotoxin may also affect the results of functional studies, we recommend the NA/LE (No Azide/Low Endotoxin) antibody format, if available, for in vitro and in vivo use.
  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. An isotype control should be used at the same concentration as the antibody of interest.
565292 Rev. 1
Antibody Details
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EMR8-5

The EMR8-5 monoclonal antibody recognizes a conserved epitope expressed on denatured human Major Histocompatibility Complex (MHC) Class I antigen heavy chain proteins, also known as, HLA (Human Leucocyte Antigens): HLA-A, HLA-B, and HLA-C (HLA-ABC). MHC Class I molecules are heterodimers that consist of a ~40-45 kDa transmembrane alpha chain subunit that is highly polymorphic and noncovalently associated with invariant β2-microglobulin. MHC Class I antigens are normally expressed on most all nucleated cells. They can be highly expressed on certain activated cells or cells treated with a variety of agents including cytokines or inflammatory mediators. In contrast, they may be very lowly expressed on certain virus-infected or tumor cells. HLA class I antigens expressed by thymic epithelial cells are involved in the positive and negative selection of CD8+ T cells and their TCR repertoire during T cell maturation within the thymus. In the periphery, these HLA antigens serve to present antigens for the generation of effector and memory CD8+ T cell responses. MHC Class I recognition by certain NK receptors can protect target cells from NK cell-mediated lysis. The EMR8-5 antibody is useful for staining HLA Class I molecules found in formalin-fixed cells or paraffin-embedded tissue sections.

565292 Rev. 1
Format Details
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Purified
Tissue culture supernatant is purified by either protein A/G or affinity purification methods. Both methods yield antibody in solution that is free of most other soluble proteins, lipids, etc. This format provides pure antibody that is suitable for a number of downstream applications including: secondary labeling for flow cytometry or microscopy, ELISA, Western blot, etc.
Purified
565292 Rev.1
Citations & References
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Development References (2)

  1. Torigoe T, Asanuma H, Nakazawa E, et al. Establishment of a monoclonal anti-pan HLA class I antibody suitable for immunostaining of formalin-fixed tissue unusually high frequency of down-regulation in breast cancer tissues. Pathol Int. 2012; 62(5):303-308. (Immunogen: ELISA, Immunohistochemistry, Western blot). View Reference
  2. Trowsdale J, Lee J, Kelly A, et al. Isolation and sequencing of a cDNA clone for a human HLA-ABC antigen. Mol Biol Cell. 1984; 2(1):53-61. (Biology). View Reference
565292 Rev. 1

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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.