Apo-ONE® Homogeneous Caspase-3/7 Assay

The Apo-ONE® Homogeneous Caspase-3/7 Assay provides the necessary reagents for fast and sensitive measurement of active caspase-3 and -7 in a homogeneous format. The assay includes a profluorescent caspase-3/7 consensus substrate, rhodamine 110 bis-(N-CBZ-l-aspartyl-l-glutamyl-l-valyl-aspartic acid amide) (Z-DEVD-R110), and an optimized bifunctional cell lysis/activity buffer. The buffer efficiently lyses cultured mammalian cells and supports optimal caspase-3/7 enzymatic activity. The substrate and buffer are combined to make the Apo-ONE® Caspase-3/7 Reagent that is added directly to samples. Upon cleavage on the C-terminal side of the aspartate residue in the DEVD peptide substrate sequence by caspase-3/7 enzymes, the rhodamine 110 becomes fluorescent when excited at a wavelength of 498nm. The emission maximum is 521nm. The amount of fluorescent product generated is representative of the amount of active caspase-3/7 present in the sample.

Features and Benefits

Get Results Faster: The simple "add-mix-measure" format combined with the high sensitivity of the assay dramatically decreases the "time to first result" by eliminating cumbersome sample preparation and lengthy incubation steps.

Use Less Enzyme or Fewer Cells: Optimized caspase-3/7 activity buffer, in conjunction with the R110-labeled substrate, allows for increased sensitivity over existing fluorescent caspase assay methods.

Adapt to Your Format and Throughput Needs: The assay can be flexibly configured (from cuvette to 384-well plate) for use in high-throughput systems by maintaining a 1:1 ratio of sample to assay reagent and may be used with purified enzyme preparations, cell extracts or cultures of adherent, suspension or primary cells.

Get More Information: Perform more than one assay on the same sample. This assay can be multiplexed with other assay methods such as the CellTiter-Blue® Assay (Cat.# G8080) or the Caspase-Glo® 8 or 9 Assays (Cat.# G8200 or G8210).

Using 100μl of Apo-ONE® Reagent per assay in a 96-well format, Cat.# G7792 provides sufficient reagents to perform 10 assays; Cat.# G7790, 100 assays; Cat.# G7791, 1,000 assays. Using 25μl of Apo-ONE® Reagent per assay in a 384-well format, Cat.# G7792 provides sufficient reagents to perform 40 assays; Cat.# G7790, 400 assays; Cat.# G7791, 4,000 assays.