Both M-calpain and mu-calpain are ubiquitously expressed, and are countered by the endogenous calpain inhibitor, calpastatin. Other calpain family members (calpain-94, ncl-2, ncl-3, etc) have more limited tissue distribution, and perhaps different functions. The calpain family members consist of a common small subunit (Calpain-4), and a large variable subunit. It is not clear that all calpains contain 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. RP2-Calpain2 recognizes latent, but not amino-processed M-calpain; the latent large subunit is 80 Kd, and the amino terminal truncation at activation yields an approximately 58 Kd form. Cleavage of the carboxyterminal region generates smaller forms of M-Calpain, but it is not clear if these forms are proteolytically active. RP2-Calpain2 Anti-M-Calpain works well for Western Blotting applications, and when used against the reduced protein identifies bands at 80 Kd, 68 Kd, and a series of further cleaved forms. RP2-Calpain2 will also bind to the non-reduced protein, and thus has application for immunoprecipitation, immunohistochemistry, and ELISA. Most cell types produce M-Calpain; since it is a cellular protein, cell lysate works well for Western blots. Rat, mouse and human M-Calpain are detected with RP2Calpain2. 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.

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