Multiparameter flow cytometric analysis of HLA-C expression on human peripheral blood leucocyte populations. The erythrocytes from human blood were lysed with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899). After washing, the remaining leucocytes were stained with either PE Mouse IgG2b, κ Isotype Control (Cat. No. 555058, Left Plot) or PE Mouse Anti-Human HLA-C antibody (Cat. No. 566372; Right Plot). Two-parameter flow cytometric contour plots showing the correlated expression of HLA-C (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 FACSCanto™ II Flow Cytometer System.
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Multiparameter flow cytometric analysis of HLA-C expression on human peripheral blood leucocyte populations. The erythrocytes from human blood were lysed with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899). After washing, the remaining leucocytes were stained with either PE Mouse IgG2b, κ Isotype Control (Cat. No. 555058, Left Plot) or PE Mouse Anti-Human HLA-C antibody (Cat. No. 566372; Right Plot). Two-parameter flow cytometric contour plots showing the correlated expression of HLA-C (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 FACSCanto™ II Flow Cytometer System.
Product Details
BD Pharmingen™
HLAC; HLC-C; HLA-JY3; MHC class I antigen C; PSORS1; HLA-E
Human (QC Testing)
Mouse IgG2b, κ
Purified Human Tamarin MHC Class I Molecules
Flow cytometry (Routinely Tested)
5 µl/test
AB_2739715
Aqueous buffered solution containing BSA, protein stabilizer, 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 with R-PE under optimum conditions, and unconjugated antibody and free PE were removed.
Product Notices
- 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).
- An isotype control should be used at the same concentration as the antibody of interest.
- Source of all serum proteins is from USDA inspected abattoirs located in the United States.
- 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.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
Companion Products
Stain Buffer (FBS) RUO
Size
500 mL
Cat No.
554656
Stain Buffer (BSA) RUO
Size
500 mL
Cat No.
554657
Lysing Buffer RUO
Size
100 mL
Cat No.
555899
Lysing Solution 10X Concentrate IVD
Size
100 mL
Cat No.
349202
PE Mouse IgG2b, κ Isotype Control RUO
Size
0.1 mg
Cat No.
555058
566372 Rev. 1
Antibody Details
DT-9
The DT-9 monoclonal antibody specifically recognizes Human Leukocyte Antigen (HLA-C), a polymorphic major histocompatibility complex (MHC) class I antigen. HLA-C is a heterodimer comprised of the HLA-C alpha chain, a ~45 kDa type I transmembrane glycoprotein, and a ~12 kDa Beta-2 (β2)-microglobulin light chain. HLA-C is expressed on nearly all cells, and plays a role in the antigen-specific, MHC-restricted presentation of small peptides to CD8+ T cells in the generation of immunity or tolerance. HLA-C may also bind to regulatory MHC class I antigen-selective receptors expressed by CD8+ T cells and natural killer (NK) cells. The DT-9 antibody also recognizes some rare HLA-A and HLA-B allotypes and the nonclassical HLA-E molecule.
566372 Rev. 1
Format Details
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.
PE
Yellow-Green 488 nm, 532 nm, 561 nm
496 nm, 566 nm
576 nm
566372 Rev.1
Citations & References
View product citations for antibody "566372" on CiteAb
Development References (6)
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Apps R, Qi Y, Carlson JM, et al. Influence of HLA-C expression level on HIV control.. Science. 2013; 340(6128):87-91. (Clone-specific: Flow cytometry). View Reference
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Braud VM, Allan DS, O'Callaghan CA, et al. HLA-E binds to natural killer cell receptors CD94/NKG2A, B and C.. Nature. 1998; 391(6669):795-9. (Clone-specific: Flow cytometry). View Reference
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Braud VM, Allan DS, Wilson D, McMichael AJ. TAP- and tapasin-dependent HLA-E surface expression correlates with the binding of an MHC class I leader peptide.. Curr Biol. 1998; 8(1):1-10. (Immunogen: Flow cytometry, Immunoprecipitation). View Reference
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Corrah TW, Goonetilleke N, Kopycinski J, et al. Reappraisal of the relationship between the HIV-1-protective single-nucleotide polymorphism 35 kilobases upstream of the HLA-C gene and surface HLA-C expression.. J Virol. 2011; 85(7):3367-74. (Clone-specific: Flow cytometry). View Reference
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Thomas R, Apps R, Qi Y, et al. HLA-C cell surface expression and control of HIV/AIDS correlate with a variant upstream of HLA-C.. Nat Genet. 2009; 41(12):1290-4. (Clone-specific: Flow cytometry). View Reference
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Zipeto D, Beretta A. HLA-C and HIV-1: friends or foes?. Retrovirology. 2012; 9:39. (Biology). View Reference
566372 Rev. 1
Please refer to Support Documents for Quality Certificates
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.