SensoLyte® Homogeneous AFC Caspase-3/7 Assay Kit *Fluorimetric*

Central to the execution phase of apoptosis are the two closely related caspase-3 and caspase-7. They share common substrate specificity and structure, but differ completely in the sequence of their respective N-terminal regions. Both caspase-3 and caspase-7 have substrate selectivity for the amino acid sequence Asp-Glu-Val-Asp (DEVD).
The SensoLyte® Homogeneous AFC Capase-3/7 Assay Kit uses Ac-DEVD-AFC as the fluorogenic indicator for assaying caspase-3/7 activities. Upon caspase-3/7 cleavage, Ac-DEVD-AFC generates the AFC fluorophore which has bright blue fluorescence and can be detected at Ex/Em=380 nm/500 nm. A bi-function assay buffer in this kit is designed to lyze the cells and measure the enzyme activity at the same time. Thus, this kit can measure caspase-3/7 activity in cell culture directly in a 96-well or 384-well plate without a time-consuming cell extraction step. In case the cells are cultured in larger plates or flasks, a lysis buffer and protocol for cell lysate preparation are also conveniently included in the kit. The kit is suitable for high throughput screening of apoptosis inducers and inhibitors.

Kits components     
Component A: Caspase-3/7 substrate, Ex/Em=380 nm/500 nm: 270 µL Component B: AFC, fluorescence reference standard, Ex/Em=380 nm/500 nm: 10 mM, 20 µL Component C: Caspase-3/7 inhibitor: 5 mM, 15 µL Component D: Assay Buffer: 30 mL Component E: DTT: 1 M, 1 mL Component F: 10X Lysis Buffer: 20 mL

Properties
Absorbance (nm): 380
Emission (nm): 500