Fluorometric assay kit for caspase-12 activity
Caspase family of proteases are the central mediators of apoptosis in mammalian cells. The Caspase-12 Fluorometric Assay Kit provides a simple and convenient means for assaying the activity of caspases that recognize the sequence ATAD. The assay is based on detection of cleavage of substrate ATAD-AFC (AFC: 7-amino-4-trifluoromethyl coumarin). ATAD-AFC emits blue light (max = 400 nm); upon cleavage of the substrate by caspase-12 or related caspases, free AFC emits a yellow-green fluorescence (max = 505 nm), which can be quantified using a fluorometer or a fluorecence microtiter plate reader.
Detect early/middle stages of apoptosis; differentiate apoptosis from necrosis.
For research use only. Not intended for any diagnostic or therapeutic purpose.
Store kit at -20°C (Store Cell Lysis Buffer & 2X Reaction Buffer at 4°C after opening). All reagents are Stable for 6 months from date of receipt under proper storage conditions.
Cell Lysis Buffer: 100ml;2X Reaction Buffer: 4*2ml; ATAD-AFC (1mM): 0.5ml; DTT (1M): 0.4ml
Fluorecence microtiter plate reader (400-nm excitation filter and 505-nm emission filter) and fluorescence microscopy
Compatible Sample Types:
Cell and tissue lysates
Features & Benefits:
1. Simple one-step procedure; takes only 1-2 hours.2. Fast and convenient.3. Comparison of the fluorescence of AFC from an apoptotic sample with an uninduced control allows determination of the fold increase in caspase-12 activity.