Calpain-6, also known as calpamodulin or calpain-X, is an intracellular, calcium-dependent 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. 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-6 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 the X chromosome, the calpain-6 sequence lacks the "EF hand" calcium-binding motif found in domain-IV of the classical calpains. 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. RP1Calpain-6 recognizes the propeptide domain of Calpain-6, which is proteolytically removed during activation. RP1Calpain-6 can be used to distinguish latent Calpain-6 from active forms; the latent large subunit is 75 kD, and the amino terminal truncation at activation yields an approximately 58kD form. A cascade of smaller forms can be seen with further activation. RP1Calpain-6 Anti-Calpain-5 works well for Western Blotting applications, and when used against the reduced protein identifies bands at 75 kD, 58kD, and a series of further cleaved active forms. RP1Calpain-6 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-6 are detected with RP1Calpain-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

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