Caspase 7 Activity Assay Kit, BioAssay™ (Mch-3, ICE-LAP3, CMH-1)
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| Caspases are cysteine protease mediators of apoptosis, destroying normal cellular function by cleaving substrates at Asp residues (1). Caspase-7 (CMH-1, Mch3, ICE-LAP3) has been identified as a major contributor to the execution of apoptosis (2–4). Caspase-7 is an effector caspase (along with Caspases-3 and 6), meaning it cleaves essential cellular machinery rather than activating other caspases (5–7). Caspase-7 is cleaved from an inactive proenzyme to an active form, a heterotetramer of two large and two small subunits. Caspase-7 is cleaved by many enzymes, including Caspases-3, 6, 8, 9 and granzyme B (1,4,5). Once activated, Caspase-7 cleaves many of the same substrates as Caspase-3, including poly (ADP-ribose) polymerase, or PARP (2,4). This kit includes all reagents required to measure Caspase-7 activity, including antibody, substrate, reaction buffer and cell extracts buffer. The assay system uses Caspase-7 antibody to selectively immunoprecipitate Caspase-7. Fluorometric substrate is added and is cleaved proportionally to the amount of activated Caspase-7, generating free fluorescent AFC. Free AFCs are determined fluorometrically at lambda max = 505nM. Compared to other commercially available Caspase-7 activity assays, this kit is designed to be more specific by using Caspase-7 antibody to selectively immunoprecipitate Caspase-7 from cell lysates. | | | Catalog # | C2087-29P | | Kit Components | C2087-29P1: Caspase 7 Pab, 1x150ul. | | Supplied as a liquid in 10mM sodium HEPES (pH 7.5), 150mM NaCl, 0.1mg/ml BSA and 50% glycerol. Purifed by immunoaffinity chromatography. Store at -20ºC. | | C2087-29P2 | Substrate (Ac-Asp-Glu-Val-Asp-AFC), 1x100ul | | C2087-29P3. Cell Extract Buffer (10X), 1x1.5ml At 1X concentration | 50mM PIPES/KOH (pH 6.5), 2mM EDTA, 0.1% CHAPS, 5mM DTT, 20µg/ml Leupeptin, 10µg/ml Pepstatin A and 10µg/ml Aprotinin. | | C2087-29P4 | Reaction/Washing Buffer (10X), 1x1ml. At 1X concentration 50mM HEPES (pH 7.5), 75mM NaCl, 5mM DTT, 0.1% CHAPS and 200X DTT (1M DTT). | | Applications | Suitable for use in Immunoprecipitation. Other applications not tested. | | Recommended Dilution | C2087-29P1-Immunoprecipitation: 1:100 | | Optimal dilutions to be determined by the researcher. | | Purity | Purified by protein A and peptide affinity chromatography. | | Form: Supplied as a liquid in 10mM sodium HEPES, pH 7.5, 150mM sodium chloride, 100 ug/ml BSA, 50% glycerol. | | Storage and Stability | May be stored at 4°C for short-term only. For long-term storage, store at -20°C. Aliquots are stable for at least 6 months at -20°C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer. | | Important Note | This product as supplied is intended for research use only, not for use in human, therapeutic or diagnostic applications without the expressed written authorization of United States Biological. |
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