Contents:

Storage

Store at 4℃.Component A should be protected from light. Expiration date marked on the outer packing.

Parameters

SuperView™ 488 Abs/Em: 500/530 nm (with DNA)

Description

This kit contains caspase-3 substrate and a caspase-3 inhibitor Ac-DEVD-CHO. It provides a convenient tool for profiling apoptotic cell population based on caspase-3 activity using fluorescence microscopy or flow cytometry.

Compared with other fluorescence substrates or inhibitors of caspase based on FLICA analysis, SuperView™ 488 Caspase-3 Substrate can detect activity of caspase-3 without inhibiting the whole cell apoptosis process.

Unlike conventional caspase assays, SuperView™ 488 Caspase-3 Substrate detects activity of caspase-3 within individual intact cells without inhibiting it. The substrate consists of a fluorogenic DNA dye and a DEVD substrate moiety specific for caspase-3. The substrate, which is both non-fluorescent and nonfunctional as a DNA dye, rapidly crosses cell membranes to enter the cytoplasm, where it is cleaved by caspase-3 to form a high-affinity DNA dye that stains the nucleus bright green. Thus, the SuperView™ 488 Caspase-3 Substrate is bi-functional, allowing detection of intracellular activity of caspase-3 and visualization of changes in nuclear morphology during apoptosis. The fluorescent staining produced in response to activity of caspase-3 is compatible with subsequent fixation and permeabilization for immunostaining.

Notes 

1. Cells can be counterstained with Hoechst 33342 at a final concentration of 1 μM to stain nucleus with blue fluorescence (Ex/Em: 346/460 nm) 

2. SuperView™ 488 staining is formaldehyde-fixable and not compatible with methanol fixation. 

3. Formaldehyde-fixed SuperView™ 488-stained cells can be permeabilized with 0.1% Triton X-100 for subsequent immunostaining; however, staining brightness may be diminished after permeabilization and washing.