Mouse anti Human HLA Class I Heavy Chain (Restricted expression)

The antibody HCA2 reacts preferentially with HLA-A locus heavy chains. HCA2 was raised against free class I heavy chains of HLA, to obtain antibodies that would still react with denatured class I antigens, as they occur in Western blotting, conventional light microscopical analysis of formalin-fixed, paraffin-embedded sections, and cryo-immuno-electron microscopy. HCA2 indeed retains strong reactivity with free class I heavy chains in Western blots. HCA2 in particular reacts in a locus-specific manner by biochemical criteria. Conditions are described for use of HCA2 in immunohistochemical staining of formalin-fixed, paraffin-embedded sections (see references). HCA2 also produces strong reactivity in immuno-electron microscopy. Its use allows the determination of tissue and subcellular distribution of class I antigens.

Optimal antibody dilutions for the different applications should be determined by titration; recommended range is 1:100 ? 1:200 for flow cytometry, and for immunohistochemistry with avidin-biotinylated horseradish peroxidase complex (ABC) as detection reagent, and 1:100 ? 1:1000 for immunoblotting applications.

The HLA class I gene family is composed of a group of genes whose products encode cell surface glycoproteins of MW 40?45 kDa, associated non-covalently with the beta-2-microglobulin light chain. They include the three polymorphic molecules HLA-A, -B, and -C, which are ubiquitously expressed and which are able to present intracellular peptides to cytotoxic T cells. Three additional class I genes are known, commonly referred to as non-classical or class Ib genes, all highly homologous to the other class I genes and all of which associate with beta-2-microglobulin light chain. In humans, each of the class Ib genes appears to exhibit a distinct pattern of expression in developing
and adult tissues. HLA-E transcripts are distributed widely in adult tissues and have also been found in the placenta and fetal liver. In the adult, the presence of HLA-F has been shown in skin, resting T cells, and B cells, whereas its expression during development has been reported in fetal liver and at low levels in placenta and extra-placental tissues. HLA-G was originally thought to be expressed only in certain populations of placental trophoblasts, but low levels have also been found in a variety of human tissues. Recently it was shown that HLA class I expression in breast cancer cells can have a predictive value for chemotherapy response.

Immunology|Inflammation

Each vial contains 100 ?l 1 mg/ml purified monoclonal antibody in PBS containing 0.09% sodium azide.

Each vial contains 100 ?l 1 mg/ml purified monoclonal antibody in PBS containing 0.09% sodium azide.

HCA2 is a mouse monoclonal IgG1 antibody derived by fusion of SP2/0-Ag14 mouse myeloma cells with spleen cells from BALB/c mice immunized with HLA-B7 and -B40 heavy chains.

The mouse monoclonal antibody HCA2 recognize HLA class I heavy chains. The reactivity spectrum of HCA2 is composed of all HLA-A chains (except HLA-A24), as well as some HLA-B, HLA-C, HLA-E, HLA-F, and HLA-G chains.