Caspases are a family of cysteine proteases that are key mediators of programmed cell death or apoptosis (1). The precursor form of all caspases is composed of a prodomain, and large and small catalytic subunits. The active forms of caspases are generated by several stimuli including ligand-receptor interactions, growth factor deprivation and inhibitors of cellular functions. All known caspases require cleavage adjacent to aspartates to liberate one large and one small subunit, which associate into a2b2 tetramer to form the active enzyme. Gene for Caspase-3 also known as Yama, CPP32, and apopain codes for a 32kD protein (2-4). Caspase-3 cleaves the death substrate poly(ADP-ribose) polymerase (PARP) to a specific 85kD form observed during apoptosis and is inhibitable by the CrmA protein. Other Caspase-3 substrates include DNA-PK, actin, GAS2, and procaspase-6, etc. (5). Caspase-3 is activated by cleavage events at Asp-28/Ser-29 (between N-terminal pro-domain) and Asp-175/Ser-176 (between large and small subunits) to generate a large subunit of 17kD and a small subunit of 12kD (3).
Positive Control: HL60 or staurosporine treated apoptotic HeLa
Applications:
Suitable for use in Western Blot. Other applications not tested.
Recommended Dilution:
Optimal dilutions to be determined by the researcher.
Storage and Stability:
May be stored at 4 degrees C for short-term only. For long-term storage and to avoid repeated freezing and thawing, aliquot Store at -20 degrees C. Aliquots are stable for at least 12 months at -20 degrees 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.
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