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| Data Sheet for RP1Calpain5 | |
| Product Name | Rabbit Antibody to Calpain-5 (H-TRA-3); Aminoterminal end Domain-I |
| Catalog Number | RP1Calpain5 |
| Product size | 100 ug |
| Description | RP1Calpain-5 is a Rabbit antibody to the cysteine protease, calpain-5, made with a synthetic peptide based on the amino terminal end of the large subunit, using the human sequence. The antibody binds to calpain-5, but does not cross react with the other calpain family members (calpain-1, calpain-2, Calpain-3, ncl-3, etc.). RP1Calpain-5 reacts to the first 30 amino acids of the "propeptide domain," (Domain-I) of Calpain-5, aminoterminal from the activation cleavage site. The antibody does not recognize amino-processed Calpain-5, 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 |
Calpain proteinases are defined currently as papain-like neutral proteases that are calcium activated. Calpains 1 and 2 are composed of a large subunit, which is proteolytically active, and a small subunit (also called calpain-4), or enhancer protein, that is not proteolytically active. The other calpain proteins identified to date are not known to require 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). Calpain-5, also known as the human orthologue of TRA-3, a C. elegans protein involved in sex determination, is located on chromosome 11 in humans, near calpain-1.Calpain-5 domain-T does not have the standard calpain EF hand domain for binding calcium, making it unclear whether it is calcium regulated. It is also unclear if calpain-5 plays any role in mammalian sex determination, since the enzyme is found in colon, kidney, liver and trachea and other tissues, as well as testis.RP1Calpain-5 recognizes latent, but not amino-processed calpain-5; the latent large subunit is 73 Kd, and the amino terminal truncation at activation yields an approximately 58 Kd form. Cleavage of the carboxyterminal region generates smaller forms of Calpain-5, but it is not clear if these forms are proteolytically active. RP1Calpain-5 works well for Western Blotting applications, and when used against the reduced protein identifies bands at 73 Kd, 68 Kd, and a series of further cleaved forms. RP1Calpain-5 will also bind to the non-reduced protein, and thus has application for immunoprecipitation, immunohistochemistry, and ELISA. Rat, mouse and human Calpain-5 are detected with RP1Calpain-5. 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. |
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