Class I Human Leukocyte Antigens (HLA -C ) are heterodimeric (heavy chain + β2m) surface molecules that bind small peptide antigens and engage T lymphocytes and NK cells expressing the T cell receptor and Killer Immunoglobulin- like Receptors (KIR).

PRODUCT

1. Each vial contains the indicated amount of IgG (caprylic acid purified) in 0.1% gelatine and 0.05% NaN3.

2. Unpurified reagent is provided at the indicated amounts with 0.1% NaN3.

Centrifuge the vial prior to use.

SPECIFICITY

Mab L31 (IgG1) binds a linear epitope on HLA class I heavy chain alleles carrying an aromatic residue (Y/F) at position 67 (ref.1,2). These include most HLA-C (HLA-Cw1 through HLA-Cw8 + others) and a few crossreacting HLA-B (HLA-B7, -B8, -B35, -B51 + others) alleles (ref.2). L31 binds when these heavy chain are free of β 2m, denatured, or unfolded (ref. 1-6).

STORAGE

Store frozen in aliquots and avoid repeated freezethawing. Stable 5 yrs.

SHIPPING CONDITIONS

Room temperature.

RESEARCH USE

This antibody is sold for laboratory research use only, not for human or in vivo use.

APPLICATIONS

Optimal working conditions in each assay may vary in different labs, and titration is advisable.

Flow cytometry, L31 (10 to 20 µg/ml) will stain ‘free’ heavy chains poorly surface-expressed in lymphoid and nonlymphoid cells and PBMCs (ref-1-3).

Immunohistochemistry (fluorescence, enzymatic), Acetone-fixed cryostatic sections and cytospins (10 to 50 µg/ml) (ref.3). Formalin-fixed, paraffin-embedded sections as described (ref.4). Under these conditions a concentration of 100 µg/ml and overnight incubation were found to provide the most consistent staining patterns.

Immunoprecipitation, One to 5 µg using rabbit anti-mouse Ig and protein ASepharose beads (ref. 2-3,5,6).

Western Blotting, Use at 10 µg/ml or less (ref.2).

REFERENCES
1. J Exp Med (1991) 174: 53-62.
2. Hum Immunol (1996) 46: 69-81.
3. Tissue Antigens (1997) 50: 555-566.
4. Am J Surg Pathol (2007) 31: 76-84.
5. J Biol Chem (2008) 283: 16469-16476.
6. J Biol Chem (2008) 283: 1267-1274.