Incucyte® Caspase-3/7 Apoptosis Stain

Incucyte® Caspase-3/7 Apoptosis Stains are formulated to detect apoptosis (cell death) in living cells. Stains were adapted for real-time quantification of cells undergoing caspase-3/7-mediated apoptosis. Addition of Incucyte® Caspase-3/7 Green, Red or Metabolism Dyes to healthy cells does not interfere with cell growth and morphology. When used with the Incucyte® real-time cell monitor software, these reagents provide reliable and accurate assessment of cell death.

Incucyte® Caspase-3/7 Apoptosis Dyes bind the activated caspase-3/7 recognition motif (DEVD) to the intercalating DNA dye. When the reagent is added to the tissue culture medium, the inert, non-fluorescent substrate passes through the cell membrane where it is cleaved by activated caspase-3/7, leading to the release of the intercalating DNA dye and fluorescent staining of DNA in the nucleus. With Incucyte® integrated analysis software, fluorescent objects can be quantified and background fluorescence minimized. These reagents have been validated for use with Incucyte® devices and allow real-time assessment of cellular apoptosis induced by pharmacological agents and/or genetic and environmental factors.

Storage and color stability

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

Recommended use

We recommend diluting Incucyte® Caspase-3/7 dyes in whole culture medium and adding them directly to cells in culture, according to the guidelines below:

• Incucyte® Caspase-3/7 Green: 1:1000 in culture medium, for a final assay concentration of 5 μM
• Incucyte® Caspase-3/7 Row: Optimize the final assay concentration by diluting the reagent 1:200 in whole medium, then make a 2-fold dilution series for final assay concentrations of 2.5, 1.25, and 0.5 µM
• Incucyte® Caspase-3/7 for metabolism: 1:1000 in culture medium, for a final assay concentration of 1 μM.

Compatible with Incucyte® SX5, Incucyte® S3 and Incucyte® SX1 Live Cell Analyzers.