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Invitrogen
HLA-ABC Monoclonal Antibody (W6/32), NovaFluor™ Yellow 590, eBioscience™
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FIGURE: 1 / 2
HLA-ABC Antibody (H020T03Y02-A) in Flow
Normal human peripheral blood cells were either left unstained (blue histogram) or stained with HLA-ABC Monoclonal Antibody, NovaFluor Yellow 590 (purple histogram). Total viable cells in the lymphocyte gate were used for analysis, as determined by LIVE/DEAD Blue (Product # L34962). Data was acquired on a 5-laser Cytek Aurora and unmixed with autofluorescence extraction.
Product Details
H020T03Y02-A
Product Specifications
Species Reactivity
Human
Host/Isotype
Mouse
/ IgG2a, kappa
Class
Monoclonal
Type
Antibody
Clone
W6/32
Conjugate
Excitation/Emission Max
563/594 nm
View spectra
Form
Liquid
Concentration
5 µL/Test
Purification
Affinity chromatography
Storage buffer
PBS, pH 7.2
Contains
0.09% sodium azide
Storage conditions
4°C, store in dark, DO NOT FREEZE!
RRID
AB_3098253 (AB_2910688)
Product Specific Information
Description
The W6/32 monoclonal antibody reacts with the human major histocompatibility complex (MHC) class I, HLA-A, B, C. MHC class I antigens associated with beta 2-microglobulin are expressed by all human nucleated cells and are central in cell-mediated immune response and tumor surveillance. W6/32 mAb recognizes a non-polymorphic epitope shared among products of the HLA-A, B, and C loci and immunoprecipitates both 43 kDa and 11-12 kDa chains. Crossreactivity is also seen in baboon, rhesus and cynomolgus monkey.
This product contains 1 vial of NovaFluor conjugate and 1 vial of CellBlox Plus Blocking Buffer.
Applications Tested
This W6/32 antibody has been pre-titrated and tested by flow cytometric analysis of normal human peripheral blood cells. This can be used at 5 µL (0.6 µg) per test. A test is defined as the amount (µg) of antibody that will stain a cell sample in a final volume of 100 µL. Cell number should be determined empirically but can range from 10^5 to 10^8 cells/test.
Master mixes
• Master mixes of NFs should be made at 2-8 °C and may be made up to 4 hours ahead of time.
• We do not recommend storing master mixes containing NovaFluor conjugates overnight or longer.
Whole Blood compatibility
• When utilizing whole blood (as opposed to density-gradient-purified PBMC), we recommend lysing red blood cells in bulk prior to staining with NovaFluor conjugates.
• See the Bulk Lysis of Human Whole Blood protocol here.
• Staining of whole blood with NovaFluor conjugates followed by lysis of red blood cells may result in higher-than-expected background staining.
Viability dye compatibility
• NovaFluor dyes are not compatible with DNA intercalating viability dyes.
• Do not use viability dyes such as propidium iodide, 7-actinomycin D (7-AAD) and DAPI. Invitrogen LIVE/DEAD Fixable Dead Cell stains are recommended for use with NovaFluor dyes.
CellBlox Plus Blocking Buffer
• This NovaFluor conjugate comes with CellBlox Plus Blocking Buffer (Cat. No. C001T03F01), essential for optimal staining.
• Use CellBlox Plus Blocking Buffer in all experiments with NovaFluor conjugates.
• Add 5 μL per sample to antibody cocktails/master mixes (regardless of how many Novafluor-conjugated antibodies are present) before combining with cells.
• CellBlox Plus Blocking Buffer is compatible with either Super Bright Complete Blocking Buffer or Brilliant Stain Buffer and can be used in antibody cocktails/master mixes with those reagents.
• For single-color controls, use 5 μL of CellBlox Plus Blocking Buffer per 100 μL of cell sample (10^3 to 10^8 cells).
NovaFluor conjugates are based on Phiton technology utilizing novel fluorophore-containing nucleic acid dye structures that allow for engineered fluorescent signatures with consideration for spillover and spread impacts. Learn more
Our internal testing shows that NovaFluor Yellow 590 non-specifically stains B cells in SJL mice. Non-specific staining has not been observed in BALB/c or C57BL/6 mice. Other strains have not been tested. See the Antibody Testing Data for an example of this strain-dependent difference.
Excitation: 563 nm; Emission: 594 nm; Laser: 561 nm (Yellow) Laser
Target Information
HLA-class I major histocompatibility (MHC) antigens are intrinsic membrane glycoproteins expressed on nucleated cells and noncovalently associated with an invariant beta2 microglobulin. They carry foreign determinants important for immune recognition by cytotoxic T cells, thus important for anti-viral and anti-tumor defense. Human HLA-class I antigens are represented by HLA-A, HLA-B and HLA-C molecules.
HLA and MHC antibodies play a significant role in Immunopeptidomics, facilitating the identification and characterization of neoantigens through high-performance liquid chromatography coupled to tandem Mass Spectrometry.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
How to use the Panel Builder
Watch the video to learn how to use the Invitrogen Flow Cytometry Panel Builder to build your next flow cytometry panel in 5 easy steps.
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Bioinformatics
Protein Aliases: A29 protein; alpha chain of MHC class II; antigen presenting molecule; class I MHC, HLA-B antigen; epididymis secretory protein Li 83; H-2K; H-2K(d); H2-K; histocompatibility antigen; HLA -B; HLA A; HLA B-40011; HLA B44; HLA C; HLA class I antigen; HLA class I antigen HLA-B; HLA class I histocompatibility antigen C alpha chain; HLA class I histocompatibility antigen Cw-1 alpha chain; HLA class I histocompatibility antigen, A alpha chain; HLA class I histocompatibility antigen, A-30 alpha chain; HLA class I histocompatibility antigen, A-33 alpha chain; HLA class I histocompatibility antigen, A-74 alpha chain; HLA class I histocompatibility antigen, B alpha chain; HLA class I histocompatibility antigen, B-47 alpha chain; HLA class I histocompatibility antigen, B-50 alpha chain; HLA class I histocompatibility antigen, B-82 alpha chain; HLA class I histocompatibility antigen, C alpha chain; HLA class I histocompatibility antigen, Cw-15 alpha chain; HLA class I histocompatibility antigen, Cw-18 alpha chain; HLA locus; HLA molecule; HLA-A; HLA-A locus alpha 2 domain; HLA-A null; HLA-Aw33.1; HLA-B; HLA-B 1502; HLA-B alpha chain; HLA-B alpha chain (B*2706); HLA-B alpha chain (B*5002); HLA-B alpha-chain; HLA-B35; HLA-B35 antigen; HLA-Bw50 antigen; HLA-C; HLA-C alpha chain; HLA-C alpha chain (HLA-Cw*15v); HLA-C alpha chain for Cw*18GB; HLA-C alpha-chain; HLA-C antigen; HLA-Cw; HLA-Cw*04GB; HLA-Cw*1702; HLA-Cw*18GB; HLA-Cw-1503; Human leukocyte antigen A; Human leukocyte antigen B; Human leukocyte antigen C; human leukocyte antigen-C alpha chain; hypothetical protein; lecocyte antigen B; leucocyte antigen A; Leukocyte Antigen; leukocyte antigen C; leukocyte antigen class I-A; leukocyte antigen class I-B; Leukoocyte Antigen; lymphocyte antigen; major histocompatability complex B antigen; major histocompatibility antigen HLA-C; MCH class I antigen; MGC184092; MGC7052; MHC 1; MHC antigen; MHC clas I antigen; MHC Class 1; MHC class 1 antigen; MHC class I; MHC class I alpha chain; MHC class I anitgen; MHC class I anti; MHC Class I antigen; MHC class I antigen A*30; MHC class I antigen A*33; MHC class I antigen A*74; MHC class I antigen B*47; MHC class I antigen B*50; MHC class I antigen B*82; MHC class I antigen Cw*15; MHC class I antigen Cw*18; MHC class I antigen GN00104; MHC class I antigen heavy chain; MHC class I antigen heavy chain HLA-C; MHC class I antigen HLA-A heavy chain; MHC class I antigen HLA-A33; MHC class I antigen HLA-B alpha chain; MHC class I antigen HLA-B heavy chain; MHC class I antigen null protein; MHC class I antigen SHCHA; MHC class I antigene; MHC class I histocompatibility antigen; MHC class I human leukocyte antigen; MHC class I molecule; MHC class I protein; MHC class II antigen; MHC class l antigen; MHC classI antigen; MHC HLA-ABC; MHC HLA-B cell surface glycoprotein; MHC HLA-B transmembrane glycoprotein; MHC HLA-B27-HS; MHC HLA-B27d; MHC HLA-B39N; MHC I HLA; MHC1; surface antigen; truncated class I antigen; truncated MHC class I antigen
Gene Aliases: AS; Aw-33; Aw-74; B-4901; B-5001; Bw-47; Bw-50; D6S204; HEL-S-83; HLA-A; HLA-A11; HLA-A33; HLA-B; HLA-B*45ZJ; HLA-B-3506; HLA-B-3905; HLA-B-5502; HLA-B-5602; HLA-B15; HLA-B39; HLA-B49; HLA-B50; HLA-B55; HLA-B59; HLA-B61; HLA-C; HLA-Cw; HLA-DQB1; HLA-DRB1; HLA-JY3; HLAA; HLAB; HLAC; HLC-C; MHC; PSORS1; SPDA1
UniProt ID: (Human) P04439, (Human) Q9GJ17, (Human) P10321
Entrez Gene ID: (Human) 3105, (Human) 3106, (Human) 3107
antigen processing and presentation of peptide antigen via MHC class I
antigen processing and presentation of exogenous peptide antigen via MHC class I, TAP-dependent
antigen processing and presentation of exogenous peptide antigen via MHC class I, TAP-independent
viral process
antigen processing and presentation
regulation of immune response
interferon-gamma-mediated signaling pathway
type I interferon signaling pathway
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