Apo3HTS

1.  Key Benefits

•  Homogenous assay for active caspase 3/7

•  Breakthrough in cell lysis buffer and preservation of caspase activity

•  Results in a no-wash, one-step assay

•  No need to wash out media from cell samples, just add the reagent directly to your experimental samples

•  Easy to Use: No need to make cell lysates or run Western blots

•  Works with suspension and adherent cells

2.  Assay Principle

Cell Technology’s APO 3/7 HTS Assay utilizes the quenched (z-DEVD)2-R110 peptide substrate for caspase 3/7 detection. The absorption and emission properties of the R110 dye are suppressed when attached to the z-DEVD peptide sequence. When R110 is cleaved away, by active caspase3/7, form the quenching DEVD sequence, the free dye excites at 488nm and emits at 515-530 nm. 

As a result of a novel and proprietary Lysis Buffer System, the APO 3/7 HTS Assay is a homogenous platform that can be utilized for high throughput fluorescence plate reader applications. The reagent is directly added to the samples thus eliminating any wash steps. 

Figure. In this figure, Jurkat cells were stimulated with various concentrations of staurosporine for 3 hours, after which caspase 3/7 activity was analyzed using the APO 3/7 HTS kit. 

Reference 

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Kumar, S. 1999. Mechanisms mediating caspase activation in cell death. Cell Death and Differ. 6: 1060-1066.

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