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| Data Sheet for RP1Calpain1 | |
| Product Name | Rabbit Antibody to Mu-Calpain (E.C. 3.4.22.17, Calpain-I, protein kinase-C activating factor); Aminoterminal end Domain-I. |
| Catalog Number | RP1Calpain1 |
| Product size | 100 ug |
| Description | RP1Calpain-1 is a Rabbit antibody to the cysteine protease, Mu-calpain, made with a synthetic peptide based on the amino terminal end of the large subunit, using the human sequence. The antibody binds to Mu-calpain, but does not cross react with the other calpain family members (M-calpain, calpain-94, ncl-2, ncl-3, etc.). RP1Calpain-1 reacts to the first 30 amino acids of the “propeptide domain,” (Domain-I) of Calpain-1, aminoterminal from the activation cleavage site. The antibody does not recognize amino-processed Calpain-1, and can be used to discriminate between latent and amino-processed forms. The antibody has been peptide-affinity purified, concentrated to 1 mg/ml, with the addition of 0.05% Sodium Azide as preservative, and 50% glycerol as cryoprotectant |
Use |
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. RP1Calpain-1 recognizes latent, but not amino-processed Mu-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 Mu-Calpain, but it is not clear if these forms are proteolytically active. RP1Calpain-1 Anti-Mu-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. RP1Calpain-1 will also bind to the non-reduced protein, and thus has application for immunoprecipitation, immunohistochemistry, and ELISA. Most cell types produce Mu-Calpain; since it is a cellular protein, cell lysate works well for Western blots. Rat, mouse and human Mu-Calpain are detected with RP1Calpain-1. 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. |
| Storage | The undiluted antibody solution is stable for approximately 6 months at 4°C. For longer storage, store at -20°C. The solution is buffered from freezing at -20°C by 50% glycerol, but will freeze at -80°C. Avoid unnecessary freeze-thaw cycles. Diluted antibody can be reused several times if not contaminated; store at 4°C with 0.05% Sodium Azide as preservative. |
Ordering |
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