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PE Mouse Anti-Human HLA-A
PE Mouse Anti-Human HLA-A

Multiparameter flow cytometric analysis of HLA-A expression on human peripheral blood leucocyte populations. Human whole blood was stained with either PE Mouse IgG1, κ Isotype Control (Cat. No. 554680; Left Plot) or PE Mouse Anti-Human HLA-A antibody (Cat. No. 567739; Right Plot). After staining, erythrocytes were lysed with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899). The bivariate pseudocolor density plot showing the correlated expression of HLA-A (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 viable leucocyte populations. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ software.

Multiparameter flow cytometric analysis of HLA-A expression on human peripheral blood leucocyte populations. Human whole blood was stained with either PE Mouse IgG1, κ Isotype Control (Cat. No. 554680; Left Plot) or PE Mouse Anti-Human HLA-A antibody (Cat. No. 567739; Right Plot). After staining, erythrocytes were lysed with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899). The bivariate pseudocolor density plot showing the correlated expression of HLA-A (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 viable leucocyte populations. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ software.

Product Details
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BD Pharmingen™
HLA-A; HLAA; MHC class I antigen HLA-A heavy chain
Human (QC Testing)
Mouse IgG1, κ
Human Peripheral Blood Lymphocytes
Flow cytometry (Routinely Tested)
5 µl
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.

Product Notices

  1. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  2. 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.
  3. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  4. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
  5. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
  6. 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).
  7. An isotype control should be used at the same concentration as the antibody of interest.
567739 Rev. 3
Antibody Details
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1082C5

The 1082C5 monoclonal antibody (also known as clone 108-2C5) recognizes a shared antigenic determinant present on the extracellular region of Human Leukocyte Antigen-A (HLA-A) heavy chains encoded by a limited number of different HLA-A alleles (HLA-A2, -A3, -A28, -A29, -A30, -A31 and -A33). HLA-A antigens belong to the major histocompatibility complex (MHC) of class I antigens along with HLA-B and HLA-C antigens. HLA class I molecules are heterodimers comprised of an ~40-45 kDa, highly polymorphic transmembrane α heavy chain that is a type I glycoprotein which is noncovalently-associated with an invariant β2-microglobulin (β2m) light chain. The N-terminal extracellular region of the HLA class I heavy chain is comprised of three domains (α1, α2, and α3). The α1 and α2 domains form a closed antigen-binding groove that accommodates 8-10 aa-peptide antigens. β2m non-covalently associates with the α3 heavy chain domain and promotes HLA class I stability. The intralocus HLA-A determinant recognized by the 108-2C5 antibody reportedly involves amino acids residues 76-80 of the HLA-A α1 domain. HLA-A antigens are normally expressed on all nucleated cells. These molecules play central roles in the MHC class I-restricted presentation and cross-presentation of antigens and the regulation of NK and T cell-mediated cytotoxicity that are involved in immune responses to pathogens and tumors as well as tissue allotransplantation.

        

567739 Rev. 3
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
567739 Rev.3
Citations & References
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Development References (6)

  1. Bardi MS, Jarduli LR, Jorge AJ, et al. HLA-A, B and DRB1 allele and haplotype frequencies in volunteer bone marrow donors from the north of Parana State. Rev Bras Hematol Hemoter. 2012; 34(1):25-30. (Biology). View Reference
  2. Guerreiro-Cacais AO, Uzunel M, Levitskaya J, Levitsky V. Inhibition of heavy chain and beta2-microglobulin synthesis as a mechanism of major histocompatibility complex class I downregulation during Epstein-Barr virus replication.. J Virol. 2007; 81(3):1390-400. (Clone-specific: Flow cytometry). View Reference
  3. Hühn MH, Hultcrantz M, Lind K, Ljunggren HG, Malmberg KJ, Flodström-Tullberg M. IFN-gamma production dominates the early human natural killer cell response to Coxsackievirus infection.. Cell Microbiol. 2008; 10(2):426-36. (Clone-specific: Flow cytometry). View Reference
  4. Llano M, Gumá M, Ortega M, Angulo A, López-Botet M. Differential effects of US2, US6 and US11 human cytomegalovirus proteins on HLA class Ia and HLA-E expression: impact on target susceptibility to NK cell subsets.. Eur J Immunol. 2003; 33(10):2744-54. (Clone-specific: Flow cytometry). View Reference
  5. Lozano F, Santos-Aguado J, Borche L, et al. Identification of the amino acid residues defining an intralocus determinant in the alpha 1 domain of HLA-A molecules.. Immunogenetics. 1989; 30(1):50-3. (Clone-specific: Blocking, Flow cytometry, Immunoprecipitation, Radioimmunoassay). View Reference
  6. Ryschich E, Cebotari O, Fabian OV, et al. Loss of heterozygosity in the HLA class I region in human pancreatic cancer.. Tissue Antigens. 2004; 64(6):696-702. (Clone-specific: Immunohistochemistry). View Reference
View All (6) View Less
567739 Rev. 3

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For Research Use Only. Not for use in diagnostic or therapeutic procedures.