The Caspase-3/7 Z-DEVD-R110 Assay Kit provides a method for detecting apoptosis by providing a simple and reliable method for assaying caspase-3/7 activity. The kit can be used to continuously measure the activity of caspase-3/7 in cell extracts and purified enzyme preparations, using a fluorometer or fluorescence microplate reader.
The rhodamine 110-derived substrate (Z-DEVD-R110) used in this assay is a non-fluorescent bisamide compound that, upon enzymatic cleavage, is converted in a two-step process to the fluorescent monoamide and then to the even more fluorescent R110 product. Both of these hydrolysis products exhibit spectral properties similar to those of fluorescein, with peak excitation and emission wavelengths of 496 nm and 520 nm, respectively.
In addition to the Z-DEVD–R110 substrate, the Caspase-3/7 Z-DEVD-R110 Assay Kit contains the reversible aldehyde inhibitor Ac-DEVD-CHO, as well as the reference standard R110. The Ac-DEVD-CHO inhibitor confirms that the fluorescence signals in both induced and control cell populations are due to the activity of caspase-3/7. The reference standard allows for quantification of the amount of R110 released in the reaction.
Biochemistry (1999) 38:13906-13911Product usage:
Bioorg Med Chem Lett (1999) 9:3231-3236
Cytometry A (2003) 55:15-23