Invitrogen™ EnzChek™ Caspase-3 Activity Assay Kits

Description

• Enable simple and reliable detection of caspase-3-activity-mediated apoptosis with the EnzChek Caspase-3 Activity Assay Kit, which takes advantage of specially designed substrates that fluoresce upon cleavage by the caspase-3 enzyme
• The EnzChek Caspase-3 Activity Assay Kit is available with either the Z-DEVD-AMC or Z-DEVD-R110 substrate, which yields a bright fluorescent product upon proteolytic cleavage by caspase 3/7
• The EnzChek Caspase-3 Activity Assay Kit enables detection of 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
• Key features of the EnzChek Caspase-3 Activity Assay Kit includes: • Fluorescent assay using standard fluorescein (FITC) excitation/emission settings • Format allows for continuous measurement of caspase-3/7 activity in cell extracts • Fluorescent and enzymatic controls included The aminomethylcoumarin (AMC)-derived substrate (Z-aspartic acid-glutamic acid-valine-aspartic acid (DEVD)-AMC) that is included in the EnzChek Caspase-3 Activity Assay Kit is weakly fluorescent in the UV range (excitation/emission maxima ˜330/390 nm), but yields a bright, blue-fluorescent product (excitation/emission maxima ˜342/441 nm) upon proteolytic cleavage
• The kit can be used to continuously measure the activity of caspase-3 and closely related proteases in cell extracts and purified enzyme preparations using a fluorometer or fluorescence microplate reader
• The rhodamine 110-derived substrate (Z-DEVD-R110) used in the EnzChek Caspase-3 Activity Assay Kit 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 substrates, the EnzChek Caspase Activity Assay Kit contains the reversible aldehyde inhibitor Ac-DEVD-CHO, as well as an AMC or R110 reference standard
• The Ac-DEVD-CHO inhibitor confirms that the fluorescence signals in both induced and control cell populations are due to the activity of the caspase-3/7 protease
• The reference standard allows for quantification of the amount of AMC or R110 released in the reaction.