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HLA-A2 Monoclonal Antibody (BB7.2), NovaFluor™ Yellow 755, eBioscience™
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FIGURE: 1 / 2
HLA-A2 Antibody (H070T03Y08-A) in Flow
Normal human peripheral blood cells were either left unstained (blue histogram) or stained with HLA-A2 Monoclonal Antibody, NovaFluor Yellow 755 (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
H070T03Y08-A
Product Specifications
Species Reactivity
Human
Host/Isotype
Mouse
/ IgG2b, kappa
Class
Monoclonal
Type
Antibody
Clone
BB7.2
Conjugate
Excitation/Emission Max
551/755 nm
View spectra
Form
Liquid
Concentration
0.6 µg/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
Product Specific Information
Description
This BB7.2 monoclonal antibody recognizes the human major histocompatibility complex (MHC) class I molecule HLA-A2, which is a member of the HLA-A family. MHC class I exists as a single alpha chain composed of three domains. This molecule associates with beta 2-microglobulin and is expressed on all nucleated cells. The BB7.2 monoclonal antibody has been reported to exhibit blocking activity, as well as recognize HLA-A28.
This product contains 1 vial of NovaFluor conjugate and 1 vial of CellBlox Plus Blocking Buffer.
Applications Tested
This BB7.2 antibody has been pre-diluted and tested by flow cytometric analysis of normal human peripheral blood cells. This may 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 (for 25 µg or 25 test products, Cat. No. C001T02F01; for 100 µg or 100 test, 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
Excitation: 551 nm; Emission: 755 nm; Laser: 561 nm (Yellow) Laser
Target Information
HLA-A belongs to the HLA class I heavy chain paralogues. This class I molecule is a heterodimer consisting of a heavy chain and a light chain (beta-2 microglobulin). The heavy chain is anchored in the membrane. Class I molecules play a central role in the immune system by presenting peptides derived from the endoplasmic reticulum lumen. They are expressed in nearly all cells. The heavy chain is approximately 45 kDa and its gene contains 8 exons. Exon 1 encodes the leader peptide, exons 2 and 3 encode the alpha1 and alpha2 domains, which both bind the peptide, exon 4 encodes the alpha3 domain, exon 5 encodes the transmembrane region, and exons 6 and 7 encode the cytoplasmic tail. Polymorphisms within exon 2 and exon 3 are responsible for the peptide binding specificity of each class one molecule. Typing for these polymorphisms is routinely done for bone marrow and kidney transplantation. Hundreds of HLA-A alleles have been described.
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: cell surface glycoprotein; integral membrane protein; DAQB-90C11.16; FLJ26655; heavy chain; HLA class I histocompatibility antigen, A alpha chain; HLA class I histocompatibility antigen, A-1 alpha chain; HLA protein allele A25; HLA-A; HLA-A 2402; HLA-A*0201 alpha chain; human leukocyte antigen; HLA-A*0204; HLA-A*0207 alpha chain; human leukocyte antigen; HLA-A-0102 allele; HLA-A24AK; HLA-A25; human leukocyte antigen; Human leukocyte antigen A; leucocyte antigen A; leukocyte antigen class I-A; major histocompatibility complex class I A; MHC 1; MHC class I antigen HLA-A heavy chain; MHC HLA-A cell surface antigen; MHC HLA-A10-alpha-2 chain precursor; MHC HLA-A2.4a chain; MHC HLA-A24 cell surface glycoprotein; MHC1; unnamed protein product
Gene Aliases: HLA-A; HLAA
UniProt ID: (Human) P04439
Entrez Gene ID: (Human) 3105
T cell mediated cytotoxicity
positive regulation of T cell mediated cytotoxicity
adaptive immune response
immune system process
T cell mediated cytotoxicity directed against tumor cell target
antigen processing and presentation of peptide antigen via MHC class I
antigen processing and presentation of endogenous peptide antigen via MHC class Ib
antigen processing and presentation of endogenous peptide antigen via MHC class I via ER pathway, TAP-dependent
antigen processing and presentation of endogenous peptide antigen via MHC class I via ER pathway, TAP-independent
peptide antigen assembly with MHC class I protein complex
positive regulation of T cell cytokine production
immune response
detection of bacterium
antibacterial humoral response
antigen processing and presentation
antigen processing and presentation of endogenous peptide antigen via MHC class I
positive regulation of interferon-gamma production
CD8-positive, alpha-beta T cell activation
protection from natural killer cell mediated cytotoxicity
antigen processing and presentation of exogenous peptide antigen via MHC class I
innate immune response
defense response to Gram-positive bacterium
T cell receptor signaling pathway
positive regulation of CD8-positive, alpha-beta T cell proliferation
positive regulation of memory T cell activation
positive regulation of CD8-positive, alpha-beta T cell activation
Disclaimer
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