Incucyte® Caspase-3/7 Dye for Apoptosis

Incucyte^® Caspase-3/7 Dyes for Apoptosis are formulated to detect apoptosis (cell death) in live cells. The dyes are ideally suited to the mix-and-read, real-time quantification of cells undergoing caspase-3/7 mediated apoptosis. Addition of the Incucyte^® Caspase-3/7 Green, Red or Metabolism Dyes to healthy cells is non-perturbing to cell growth and morphology. When used with Incucyte^® Live-Cell Analysis Systems and software, these reagents allow for reliable, accurate evaluation of cell death. Incucyte^® Caspase-3/7 Dyes for Apoptosis couple the activated caspase-3/7 recognition motif (DEVD) to a DNA intercalating dye. When added to tissue culture medium, the inert, non-fluorescent substrate crosses the cell membrane where it is cleaved by activated caspase-3/7, resulting in the release of the DNA dye and fluorescent staining of the nuclear DNA. With Incucyte^® integrated analysis software, fluorescent objects can be quantified and background fluorescence minimized. These reagents have been validated for use with the Incucyte^® Live-Cell Analysis System and enable real-time evaluation of cell apoptosis induced by pharmacological agents and/or genetic and environmental factors.

Presentation, Storage and Stability

The dyes are supplied as single-vial solutions in dimethyl sulfoxide (DMSO). Each vial provides a sufficient quantity for performing 100-200 tests (one test is one well of a 96-well microtiter plate). Upon receipt, the Incucyte^® Caspase 3/7 Dyes should be stored at 4°C where they are stable for six months.

Recommended Use

We recommend diluting Incucyte^® Caspase-3/7 Dyes in complete culture media and adding directly to cells in culture, as per the guidelines below:

• Incucyte^® Caspase-3/7 Green: 1:1000 in culture media, for a final assay concentration of 5 μM
• Incucyte^® Caspase-3/7 Red: Optimize final assay concentration by diluting the reagent 1:200 in complete medium, then make a series of 2-fold dilutions for final assay concentrations of 2.5, 1.25 and 0.5 µM
• Incucyte^® Caspase-3/7 for Metabolism: 1:1000 in culture media, for a final assay concentration of 1 μM

Additional protocol details can be found in the Product Guide under the Documents tab.