Use

Calpain-5, also known as the human orthologue of TRA-3, is an intracellular, calcium-dependent cysteine protease. TRA-3 is involved in sex determination in C. elegans, but the function in mammalian tissues is unknown. Calpain-5 has a more tissue-specific expression than the ubiquitous calpain-1 and calpain-2, and is found in testis, liver, kidney, trachea, and other tissues. The classical calpain family members consist of a common small subunit (calpain-4), and a large variable subunit, but it is not known if calpain-5 associates with a small subunit. 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). Located on chromosome 11 in humans, near calpain-1, the calpain-5 sequence lacks the "EF hand" calcium-binding motif found in domain-IV of the classical calpains. RP3Calpain-5 recognizes latent and active Calpain-5; the latent large subunit is 73kD, and the amino terminal truncation at activation yields an approximately 58kD form. A cascade of smaller forms can be seen with further activation. RP3Calpain-5 Anti-Calpain-5 works well for Western Blotting applications, and when used against the reduced protein identifies bands at 73kD, 58kD, and a series of further cleaved active forms. RP3Calpain-5 will also bind to the non-reduced protein, and thus has application for immunoprecipitation, immunohistochemistry, and ELISA. Calpains are cellular proteins, and cell lysate works well for Western blots. Rat, mouse and human Calpain-5 are detected with RP3Calpain-5. 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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