HLA-ABC Monoclonal Antibody (W6/32), Brilliant Ultra Violet™ 737, eBioscience™

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.

Applications Tested
This W6/32 antibody has been pre-diluted and tested by flow cytometric analysis of normal human peripheral blood cells. This may be used at 5 µL (0.5 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.

Blocking Buffers
When using two or more Super Bright, Brilliant Violet™, Brilliant Ultra Violet™, or other polymer dye-conjugated antibodies in a staining panel, it is recommended to use Super Bright Complete Staining Buffer (Product # SB-4401) or Brilliant Stain Buffer (Product # 00-4409-75) to minimize any non-specific polymer interactions. Please refer to the datasheet for Super Bright Staining Buffer or Brilliant Stain Buffer for more information.

Light sensitivity
This tandem dye is sensitive to photo-induced oxidation. Please protect this vial and stained samples from light.

Fixation
• Samples can be stored in IC Fixation Buffer (Product # 00-8222) (100 µL of cell sample + 100 µL of IC Fixation Buffer) or 1-step Fix/Lyse Solution (Product # 00-5333) for up to 3 days in the dark at 4°C with minimal impact on brightness and FRET efficiency/compensation.
• Some generalizations regarding fluorophore performance after fixation can be made, but clone specific performance should be determined empirically.
• Our internal testing suggests that Brilliant Ultra Violet™ 737 (BUV737) is compatible with short-term methanol-based fixation, but should not be stored in buffers containing methanol for longer than one hour.

Excitation: 355 nm; Emission: 732 nm; Laser: Ultraviolet Laser.

BRILLIANT ULTRA VIOLET™ is a trademark or registered trademark of Becton, Dickinson and Company or its affiliates, and is used under license. Powered by Sirigen™.

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.