Activation of ICE-family proteases/caspases initiates apoptosis in mammalian cells. The Caspase-9 Fluorometric Assay Kit provides a simple and convenient means for assaying the activity of caspases that recognize the sequence LEHD. The assay is based on detection of cleavage of substrate LEHD-AFC (AFC: 7-amino-4-trifluoromethyl coumarin). LEHD-AFC emits blue light (^max 400nm); upon cleavage of the substrate by caspase-9 or related caspases, free AFC emits a yellow-green fluorescence (^max 505nm), which can be quantified using a fluorometer or fluorecence microtiter plate reader.
Components: 25 assays 100 assays
Cell Lysis Buffer 25ml 100ml
2X Reaction Buffer 2ml 4 x 2ml
LEHD-AFC Substrate (1mM) 125ul 500ul
DTT (1M) 100ul 400ul
Stability:
Store kit at -20 degrees C (Store Cell Lysis Buffer and 2X Reaction Buffer at 4 degrees C after opening). All reagents are stable for 1 year under proper storage conditions.
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