Use

Calpain-6, most similar to Calpain-5 (the human TRA-3 ortholog), is an intracellular, papain-like cysteine protease. Calpain-6 has a much more tissue-specific expression in adults than the ubiquitous calpain-1 and calpain-2, and has thus far been found primarily in the placenta, although it is expressed embryonically in a number of tissues. In C. elegans, H-TRA-3 is involved in sex determination; the Calpain-6 gene is located on the X-chromosome, but the mammalian role is currently unknown. Domains in the large subunit include the amino terminal domain-I, the proteinase domain-II, domain-III, and the EF-hand domain-IV (domain-T in calpains 5 & 6). Calpain-6 does not have the classical EF hand calcium-binding domain-IV, and it is not clear if calcium enhances Calpain-6. In addition, the canonical active site Cys, His and Asn are modified to Lys, His, Asn in human and Lys, Tyr, Asn in mouse calpain-6, making it unlikely that calpain-6 is proteolytically active. The calpain family members consist of a common small subunit (Calpain-4), and a large variable subunit (although it is not clear if all calpains contain a small subunit). Calpain-6 has not yet been reported to associate with a small subunit. RP3Calpain-6 recognizes latent and active Calpain-6; the latent large subunit is 75 KD, and the amino terminal truncation at activation yields an approximately 58 KD form. A cascade of smaller forms can be seen with further activation. RP3Calpain-6 Anti-Calpain-6 works well for Western Blotting applications, and when used against the reduced protein identifies bands at 75 KD, 58 KD, and a series of further cleaved active forms. RP3Calpain-6 will also bind to the non-reduced protein, and thus has application for immunoprecipitation, immunohistochemistry, and ELISA. Calpain-6 is a cellular protein, and cell lysate works well for Western blots. Rat, mouse and human Calpain-6 are detected with RP3Calpain-6. A recommended starting concentration for Western blots is 1:1000 when using colorimetric substrates such as BCIP/NBT, and 1:5000 for chemiluminescent substrates. Higher concentrations of antibody may be needed for samples from more distantly related species

FOR RESEARCH USE ONLY. NOT FOR USE IN HUMANS.

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