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Invitrogen
HLA-A3 Monoclonal Antibody (GAP.A3), NovaFluor™ Blue 510, eBioscience™
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FIGURE: 1 / 2
HLA-A3 Antibody (H071T03B01-A) in Flow
Normal human peripheral blood cells were either left unstained (blue histogram) or stained with HLA-A3 Monoclonal Antibody, NovaFluor Blue 510 (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
H071T03B01-A
Product Specifications
Species Reactivity
Human
Host/Isotype
Mouse
/ IgG2a, kappa
Class
Monoclonal
Type
Antibody
Clone
GAP.A3
Conjugate
Excitation/Emission Max
493/513 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
The monoclonal antibody GAP.A3 reacts with the human major histocompatibility complex (MHC) class I molecule HLA-A3, which is a member of the HLA-A family. HLA-A3 is expressed by 15-25% of cancer patients and has been demonstrated to bind the tumor-associated antigen human telomerase reverse transcriptase (hTERT).
This product contains 1 vial of NovaFluor conjugate and 1 vial of CellBlox Plus Blocking Buffer.
Applications Tested
This GAP.A3 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 (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: 493 nm; Emission: 513 nm; Laser: 488 nm (Blue) Laser
Target Information
HLA-A3 is a subtype of the HLA class I heavy chain paralogues, existing in two forms: HLA-A3.1 and HLA-A3.2, which differ at amino acid positions 152 and 156. HLA class I molecules are heterodimers consisting of a heavy chain and a light chain (beta-2 microglobulin), with the heavy chain anchored in the membrane. These molecules are highly polymorphic cell surface glycoproteins expressed on nearly all nucleated cells. HLA class I molecules play a central role in the immune system by presenting intracellular peptides derived from the endoplasmic reticulum lumen on the cell surface. Non-native peptides are recognized by circulating cytotoxic T lymphocytes, facilitating immune surveillance. The heavy chain of HLA-A is approximately 45 kDa and its gene contains 8 exons. Exons 2 and 3 encode the alpha1 and alpha2 domains, which bind the peptide, while exon 4 encodes the alpha3 domain, exon 5 encodes the transmembrane region, and exons 6 and 7 encode the cytoplasmic tail. Polymorphisms within exons 2 and 3 determine the peptide binding specificity of each class I molecule, and typing for these polymorphisms is routinely performed for bone marrow and kidney transplantation. Hundreds of HLA-A alleles have been described, and HLA and MHC antibodies are crucial in immunopeptidomics, aiding in the identification and characterization of neoantigens through techniques such as 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; antigen presenting molecule; HLA A; HLA class I antigen; 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 locus; HLA-A; HLA-A locus alpha 2 domain; HLA-A null; HLA-Aw33.1; Human leukocyte antigen A; leukocyte antigen class I-A; MCH class I antigen; MHC class 1 antigen; 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 heavy chain; MHC class I antigen HLA-A heavy chain; MHC class I antigen HLA-A33; MHC class I antigen null protein; MHC class I antigene; MHC class II antigen; MHC classI antigen; truncated MHC class I antigen
Gene Aliases: Aw-33; Aw-74; HLA-A; HLA-A11; HLA-A33; HLA-DQB1; HLA-DRB1; HLAA
UniProt ID: (Human) Q95380
Entrez Gene ID: (Human) 3105
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
Performance Guarantee
If an Invitrogen™ antibody doesn't perform as described on our website or datasheet,we'll replace the product at no cost to you, or provide you with a credit for a future purchase.*
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