Description

Description: 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.

Additional Text

Antibody Clonality

Monoclonal

Storage Note

The antibody is shipped at ambient temperature and may be stored at +4°C. For prolonged storage prepare appropriate aliquots and store at or below -20°C. Prior to use, an aliquot is thawed slowly in the dark at ambient temperature, spun down again and used to prepare working dilutions by adding sterile phosphate buffered saline (PBS, pH 7.2). Repeated thawing and freezing should be avoided. Working dilutions should be stored at +4°C, not refrozen, and preferably used the same day. If a slight precipitation occurs upon storage, this should be removed by centrifugation. It will not affect the performance or the concentration of the product.

Application Notes

HC10 was raised against free class I heavy chains of HLA antigens 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. HC10 indeed retains strong reactivity with free class I heavy chains in Western blots. HC10 also produces strong reactivity in immuno-electron microscopy. Its use allows the determination of tissue and subcellular distribution of class I antigen and can be used to specifically block free heavy chains (FHC); See reference 6. 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.