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 AnaRed Capase-3/7 Assay Kit uses Ac-DEVD-AnaRed as the fluorogenic indicator for assaying caspase-3/7 activities. Upon caspase-3/7 cleavage, Ac-DEVD-AnaRed generates the AnaRed fluorophore has bright red fluorescence and can be detected at Ex/Em=537 nm/611 nm. The long excitation and emission wavelength of AnaRed™ is useful when apoptotic inducing compounds themselves have strong fluorescence at lower wavelength. 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.