Use

Calpain-9, also known as GC36 and nCL-4 (novel calpain #4) is an intracellular, calcium-dependent cysteine protease. Calpain-9 was first described in the digestive tract, and a shorter form with a deletion in the catalytic domain has been associated with gastric cancer. Recombinant calpain-9 has been expressed, and is unstable unless expressed with the calpain small subunit (calpain S1), suggesting that calpain-9 associates with the small subunit in-vivo. The recombinant calpain-9 showed caseinolytic activity, albeit at a lower level than native calpain-1 or calpain-2, and a calcium sensitivity intermediate between calpain-1 and calpain-2. Calpastatin blocked the caseinolytic activity. Domains in the large subunit include the amino terminal domain-I, the proteinase domain-II, domain-III, and the EF-hand domain-IV. RP4Calpain-9 recognizes latent and aminotruncated (active) calpain-9, but not the truncated form lacking the domain-IV. The latent large subunit migrates at about 79 kD on reduced SDS PAGE gels, and truncation at the amino terminal end yields an approximately 58 Kd form. A cascade of smaller forms can be seen with further activation. RP4Calpain-9 Anti Calpain-9 works well for Western Blotting applications, and when used against the reduced protein identifies bands at 80 kD, and a series of further cleaved forms. RP4Calpain-9 will also bind to the non-reduced protein, and thus has application for immunoprecipitation, immunohistochemistry, and ELISA. Not all cell types produce Calpain-9; since it is a cellular protein, cell lysate works well for Western blots. Rat, mouse and human Calpain-9 are detected with RP4Calpain-9. 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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