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| Data Sheet for RP5Calpain2 | |
| Product Name | Rabbit Antibody to M-Calpain (E.C. 3.4.22.17, Calpain-II, protein kinase-C activating factor); calcium-binding region Domain-III, large subunit |
| Catalog Number | RP5Calpain2 |
| Product size | 100 ug |
| Description | RP5calpain-2 is a Rabbit antibody to the cysteine protease, calpain-2, made with a synthetic peptide based on the calcium binding region of domain-III in the large subunit, using the human sequence. The antibody binds to M-calpain, but does not cross react with the other calpain family members (m-calpain, calpain-94, ncl-2, ncl-3, etc.). 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
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Calpain-2, also known as Calpain-M is an intracellular, calcium-dependent cysteine protease. Calpain-2 has a Millimolar sensitivity to calcium, 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 "classical" calpain family members consist of a common small subunit (calpain S1, small subunit #1), and a large variable subunit. Domains in the large subunit include the amino terminal domain-I, the proteinase domain-II, domain-III, and the EF-hand domain-IV. RP5Calpain2 recognizes latent and active M-calpain. The latent large subunit migrates at about 80 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. RP5Calpain-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. RP5Calpain-2 will also bind to the non-reduced protein, and thus has application for immunoprecipitation, immunohistochemistry, and ELISA. Most cell types produce Calpain-2; since it is a cellular protein, cell lysate works well for Western blots. Rat, mouse and human Calpain-2 are detected with RP5Calpain-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. |
| 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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