The antibody was purified by protein-G affinity chromatography, and conjugated with biotin under optimal conditions. The solution is free of unconjugated biotin.
Concentration:
0.5 mg/ml
Storage & Handling:
The antibody solution should be stored undiluted at 4°C. Do not freeze.
Each lot of this antibody is quality control tested by Western blotting. Western blotting, suggested working dilution(s): Use 5 μg antibody per 5 ml antibody dilution buffer for each mini-gel. It is recommended that the reagent be titrated for optimal performance for each application.
Jurkat cell extract was resolved by electrophoresis, transferred to nitrocellulose and probed with either purified monoclonal anti-caspase 3 antibody (lane 1) or biotinylated anti-caspase 3 (lane 2). For purified antibody, proteins were visualized using a goat anti-mouse secondary antibody conjugated to HRP and a chemiluminescence system. For biotinylated antibody, proteins were visualized using streptavidin-HRP and a chemiluminescence system. The biotinylated antibody shows several non-specific bands in Western blot that are not seen with the purified antibody (presumably proteins such as biotin-dependent carboxylases).
Description:
Caspase 3 (also known as cpp32, Apopain, Yama, and PARP cleavage protease) is a member of the peptidase family C14. This cytoplasmic caspase functions as a heterotetramer and has two isoforms (one acts as a dominant negative inhibitor). The pro-form of this caspase is approximately 32 kD. Caspase 3 is involved in the caspase activation cascade responsible for apoptosis execution. This caspase cleaves/activates SREBPs, Caspase 6, Caspase 7, and Caspase 9. Caspase 3 is inhibited by c-IAP1, c-IAP2, XIAP, survivin, and Livin and activated by Caspase 6, -8 , and -10, granzyme B, Apaf-1 by cleavage into large p17 and small p12 subunits. Active heterodimers form between the large p17 subunit and small p11 subunit of Caspase 7 and vice versa. Caspase 3 has been shown to cleave PARP, huntingtin, androgen receptor, atrophin-1, DNA- PKcs, D4-GDI, hnRNPs C1/C2, U1snRNP, PKC delta, ATM, fodrin, gelsolin, ICAD, RIP, X-IAP, STAT1, topoisomerase I, vimentin, Rb, lamin B, mdm2, and β-catenin. The 4-1-18 monoclonal antibody has been shown to be useful for Western blotting of human caspase 3 (32 kD pro-caspase and 17 kD cleaved caspase).
Peptidase family C14, heterotetramer. Two isoforms, pro-Casp3 32 kD
Distribution:
Cytoplasm
Function:
Involved in caspase activation cascade responsible for apoptosis execution. Cleaves/activates SREBPs, Casp6, Casp7, Casp9
Regulation:
Inhibited by dominant negative isoform, c-IAP1, c-IAP2, XIAP, survivin, Livin. Activated when cleaved by Casp6, -8, -10, granzyme B, Apaf-1 into large p17 and small p12 subunits
Interaction:
Active heterodimers between large p17 subunit and small p11 subunit of Casp7 and vice versa. Cleaves PARP, huntingtin, androgen receptor, atrophin-1, DNA- PKcs, D4-GDI, hnRNPs C1/C2, U1snRNP, PKC delta, ATM, fodrin, gelsolin, ICAD, RIP, X-IAP, STAT1, topo
Antigen References:
1. Goldberg, Y., et al., 1996. Nat. Genet. 13:442. 2. Fernandes-Alnemri, T., et al., 1996. PNAS 93:7464. 3. Slee, E., et al., 2001. J. Biol. Chem. 276:7320. 4. Woo, M., et al., 2003. Nat. Immunol. 4:1016.
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