Caspase 3 Apoptosis Detection, Fluorometric BioAssay(TM) Kit (CPP-32, Apoptain, Yama, SCA-1)

Activation of ICE-family proteases/caspases initiates apoptosis in mammalian cells. The Caspase-3 Fluorometric Protease Assay Kits provide a simple and convenient means for assaying the activity of caspases that recognize the sequence DEVD. The assay is based on detection of cleavage of substrate DEVD-AFC (AFC: 7-amino-4-trifluoromethyl coumarin). DEVD-AFC emits blue light (lmax 400nm); upon cleavage of the substrate by CPP32 or related caspases, free AFC emits a yellow-green fluorescence (lmax 505nm), which can be quantified using a fluorometer or a fluorescence microtiter plate reader. Comparison of the fluorescence of AFC from an apoptotic sample with an uninduced control allows determination of the fold increase in CPP32 activity. \n\nKit Components: 25 assays 100 assays\nC2087-10A Cell Lysis Buffer 25ml 100ml\nC2087-10B 2X Reaction Buffer 2ml 4 x 2ml\nC2087-10C DEVD-AFC Substrate (1mM) 125ul 500ul\nC2087-10D DTT (1M) 100ul 400ul\n\nStorage and Stability:\nStore kit at -20 degrees C (Store Cell Lysis Buffer and 2X Reaction Buffer at 4 degrees C after opening). All reagents are stable for 6 months under proper storage conditions. Protect DEVD-AFC from light.

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SPECIFICATIONS

Catalog Number

C2087-10

Size

25Tests

References

1. Brian S., et al. (2002) Pharmacol & Exp Therapeut. 302:8-17. 2. Glantschnig H., et al. (2002) J. Biol. Chem. 277:42987-42996. 3. Nelson JM and FRY DW (2001) J. Biol. Chem. 276:14842-14847. 4. Nowak, G., et al. (2002) J. Biol. Chem. 277:43377-43388. 5. Thornberry, N.A., et al., J. Biol. Chem. 272:17907-17911 (1997). 6. Casciola-Rosen, L.A., et al., J. Exp. Med., 183:1957-1964 (1996). 7. Lazebnik, Y., et al., Nature, 371:346-347 (1994)