Thermo Fisher Scientific HLA-A3 Monoclonal Antibody (GAP.A3), NovaFluor Blue 585, eBioscience

Applications and Tested Dilution

• Application: Flow Cytometry (Flow)
• Tested Dilution: 5 µL (0.6 µg)/test

Product Specifications

Additional Formats Available

• APC
• FITC
• NovaFluor Blue 510
• NovaFluor Blue 610-70S
• NovaFluor Blue 660-120S
• NovaFluor Blue 690
• NovaFluor Red 710
• NovaFluor Red 725
• NovaFluor Yellow 570
• NovaFluor Yellow 660
• NovaFluor Yellow 730
• NovaFluor Yellow 755
• PE
• Super Bright 436
• Super Bright 600
• Custom conjugation available

Product Specific Information

Description:
The monoclonal antibody GAP.A3 reacts with the human major histocompatibility complex (MHC) class I molecule HLA-A3, a member of the HLA-A family. HLA-A3 exists as two subtypes (HLA-A3.1 and HLA-A3.2) differing at amino acid positions 152 and 156. MHC class I molecules are expressed on all nucleated cells and are highly polymorphic surface glycoproteins that associate with β2 microglobulin. They present intracellular peptides to cytotoxic T lymphocytes.

HLA-A3 is expressed in 15–25% of cancer patients and binds the tumor-associated antigen human telomerase reverse transcriptase (hTERT).

Each product includes:

• 1 vial of NovaFluor conjugate
• 1 vial of CellBlox Plus Blocking Buffer

Applications Reported: Flow cytometric analysis.
Applications Tested: Pre-titrated and validated on normal human peripheral blood cells at 5 µL (0.6 µg) per test (defined for 100 µL final volume). Cell number range: 10⁵–10⁸ cells/test.

Important Usage Notes:

• NovaFluor dyes are not compatible with DNA intercalating viability dyes (e.g., propidium iodide, 7-AAD, DAPI).
• Use Invitrogen LIVE/DEAD Fixable Dead Cell stains for viability analysis.
• Use CellBlox Plus Blocking Buffer (Cat. No. C001T06F01) for optimal staining. Add 5 µL per sample before combining with cells.
• For single-color controls, use 5 µL of blocking buffer per 100 µL of cell sample (10³–10⁸ cells).

Technology:
NovaFluor conjugates utilize Phiton™ technology, employing nucleic acid dye structures engineered for optimized fluorescent signatures with reduced spillover and spread.

Excitation: 494 nm
Emission: 585 nm
Laser: 488 nm (Blue)

Target Information

HLA-A belongs to the HLA class I heavy chain paralogues. This molecule is a heterodimer with a heavy chain and β2-microglobulin light chain. The heavy chain (~45 kDa) is membrane-anchored and encoded by a gene with 8 exons:

• Exon 1: Leader peptide
• Exons 2–3: α1 and α2 domains (peptide-binding)
• Exon 4: α3 domain
• Exon 5: Transmembrane region
• Exons 6–7: Cytoplasmic tail

Polymorphisms in exons 2 and 3 determine peptide-binding specificity. HLA typing for these polymorphisms is routinely performed for bone marrow and kidney transplantation. Hundreds of HLA-A alleles have been identified.

HLA and MHC antibodies are key tools in Immunopeptidomics, aiding in neoantigen identification and characterization via HPLC-MS/MS.

For Research Use Only.
Not for use in diagnostic procedures. Not for resale without authorization.

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