Swiss-Prot link Q14790: Caspase-8 (human) (precursor)AfCS Signalling Gateway link A000503: Caspase-8 (mouse)
Swiss-Prot link P55211: Caspase-9 (human) (precursor)AfCS Signalling Gateway link A000504: Caspase-9 (mouse)
Enzyme Evaluation Protocol This protocol is used to measure caspase enzyme activity. The synthetic fluorogenic peptide Ac-IETD-AMC (Prod. No. ALX-260-042) is used as the caspase enzyme substrate. The enzyme cleaves the substrate releasing the fluorescent AMC (7-amino-4-methylcoumarin). AMC release can be measured by spectrofluorometry using UV. 1. Add 20mM of Ac-IETD-AMC (Prod. No. ALX-260-042) to 1ml of assay buffer (20mM PIPES, 100mM sodium chloride, 10mM DTT, 1mM EDTA, 0.1% (w/v) CHAPS, 10% sucrose, pH 7.2). Add the appropriate amount of active caspase to the mixture (50ng/ml).2. Incubate for 1 hour at 37°C.3. Measure the AMC liberated from the Ac-IETD-AMC using a spectrofluorometer with an excitation wavelength of 380nm and an emission wavelength of 440nm.Note: Enzyme activity can also be measured using the caspase enzyme substrate Ac-IETD-pNA (Prod. No. ALX-260-045) at 37°C in a spectromax at 405nm wavelength.Swiss-Prot link Q92851: Caspase-10 (human) (precursor)AfCS Signalling Gateway link A000493: Caspase-10 (human)