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Multiparameter flow cytometric analysis of HLA-A expression on human peripheral blood leucocyte populations. Human whole blood was stained with BD Horizon™ R718 Mouse IgG1, κ Isotype Control (Cat. No. 566928; Left Plot) or BD Horizon™ R718 Mouse Anti-Human HLA-A antibody (Cat. No. 568030/568031; Right Plot). Erythrocytes were lysed with BD FACS Lysing Solution (Cat. No. 349202). 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 intact leucocyte populations. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ software.
BD Horizon™ R718 Mouse Anti-Human HLA-A
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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.
Development References (6)
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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
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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
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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
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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
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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
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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
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