Use

M-Calpain, also known as Calpain-II, is an intracellular, calcium-dependent cysteine protease. M-calpain has a Millimolar sensitivity (thus the M-) as compared to the micromolar calcium sensitivity of m-calpain. The calpains have papain-like activity, thus the –pain nomenclature. Both M-calpain and m-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 (although it is not clear if 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. RP4Calpain-2 recognizes latent and active M-calpain; the latent large subunit is 80 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. RP4Calpain-2 Anti-M-Calpain works well for Western Blotting applications, and when used against the reduced protein identifies bands at 80 Kd, 58 Kd, and a series of further cleaved active forms. RP4Calpain-2 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 RP4Calpain-2. 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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