Application

The assay can be performed in 1 mL volume and measured using a spectrophotometer or in 100 ìl volume in a 96 well plate using an ELISA reader.

General description

Caspase 3 Colorimetric Assay Kit provides the reagents needed for a quick and efficient detection of caspase 3 activity in cell lysates and in purified preparations of caspase 3.The Caspase 3 Colorimetric Assay Kit is based on the hydrolysis of acetyl-Asp-Glu-Val-Asp p-nitroanilide (Ac-DEVD-pNA) by caspase 3, resulting in the release of the p-nitroaniline (pNA) moiety. p-Nitroaniline is detected at 405 nm (ε_mM=10.5). The concentration of the pNA released from the substrate is calculated from either the absorbance values at 405 nm or from a calibration curve prepared with pNA standards (pNA standard included with the kit).

Other Notes

Use 17 megohm water only

Unit Definition

One unit of caspase 3 will cleave 1.0 µmol of Ac-DEVD-pNA per min at pH 7.4 at 25 °C.