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...

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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 - 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).
• Automate This Assay: Validated automated methods available at: www.promega.com/automethods/
• Choose Your Configuration: Learn more about our custom options for this product at: www.promega.com/myway/

Applications

For more information, see the Protocols & Applications Guide.

Notes

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.

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