Description
Key facts
Detection method:Fluorescent,
Sample types:Tissue Extracts, Cell Lysate,
Assay type:Enzyme activity,
Assay time:2h,
Assay Platform:Microplate reader
Product details:
Caspase-3 Assay Kit (Fluorometric) (ab39383) provides a simple and convenient means for assaying DEVD-dependent caspase activity.
Note: Due to the nature of the substrate, this assay also detects caspase-7 activity.
The Caspase-3 assay protocol is based on detection of cleavage of substrate DEVD-AFC (AFC: 7-amino-4-trifluoromethyl coumarin).
DEVD-AFC emits blue light (λ max = 400 nm). On cleavage of the substrate by CPP32 or related caspases, free AFC emits a yellow-green fluorescence (Ex/Em = 400/505 nm).
The signal 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 Caspase-3/CPP32 activity.
Caspase-3 assay protocol summary:
- lyse cells / homogenize and lyse tissues in lysis buffer
- incubate on ice for 10 min
- add reaction buffer and DEVD-AFC substrate and incubate for 1-2 hr at 37°C
- analyze with fluorometer or microplate reader
REACH authorisation
Abcam has not and does not intend to apply for the REACH Authorisation of customers' uses of products that contain European Authorisation list (Annex XIV) substances.
It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses.
Properties and Storage Information:
Shipped at conditions-Blue Ice, Appropriate short-term storage conditions--20°C, Appropriate long-term storage conditions--20°C, Storage information--20°C
