How do you check caspase activity?

How do you check caspase activity?

Caspase activity was determined by quantifying fluorescence images after excitation at 380 nm and emission centered at both 545 nm (yellow fluorescence, i.e. FRET) and 450 nm (blue fluorescence, i.e. conventional excitation) were captured.

What is cell event?

CellEvent™ Caspase-3/7 Green Detection Reagent is a novel fluorogenic substrate for activated caspase-3/7 that is compatible with both live cell and fixed imaging, with absorption/emission maxima of ∼502/530 nm.

How does caspase-3/7 assay work?

Adding the single Caspase-Glo® 3/7 Reagent in an “add-mix-measure” format results in cell lysis, followed by caspase cleavage of the substrate. This liberates free aminoluciferin, which is consumed by the luciferase, generating a “glow-type” luminescent signal that is proportional to caspase-3/7 activity.

How do you use caspase 3 7 detection reagent?

To use CellEvent Caspase 3/7 Detection Reagent, simply add it to cells, incubate 30 minutes, and visualize (see figure). Apoptotic cells with activated caspase-3/7 will have bright green nuclei, while cells without activated caspase 3/7 will have minimal fluorescent signal (see figure).

What is the fluorescent emission of caspase 3/7?

The fluorescent emission of the dye when bound to DNA is ∼530 nm and can be observed using a standard FITC filter set. To use CellEvent Caspase 3/7 Detection Reagent, simply add it to cells, incubate 30 minutes, and visualize (see figure).

How do you know if caspase 3/7 is active apoptotic?

Apoptotic cells with activated caspase-3/7 will have bright green nuclei, while cells without activated caspase 3/7 will have minimal fluorescent signal (see figure). This robust assay enables the examination of caspase-3/7 activation in live cells.

Does caspase-3/7 bind to DNA?

However, after activation of caspase-3/7 in apoptotic cells, the DEVD peptide is cleaved, enabling the dye to bind to DNA and produce a bright, fluorogenic response. The fluorescent emission of the dye when bound to DNA is ∼530 nm and can be observed using a standard FITC filter set.