Method of Analysis:Flow Cytometry, Fluorescence Microscope
Intended Use:Detect active caspase enzymes with the FLICA 660 Poly Caspase BioAssay™ Kit. This in vitro assay employs the fluorescent inhibitor probe 660-VAD-FMK to label active caspase enzymes in living cells. Analyze the fluorescent signal using fluorescence microscopy or flow cytometry.
Test Principle:Caspases play important roles in apoptosis and inflammation. FLICA BioAssay™ kits are used by researchers seeking to quantitate apoptosis via caspase activity in cultured cells. The FLICA 660 Poly Caspase probe allows researchers to assess caspase activation.
The FLICA reagent 660-VAD-FMK enters each cell and irreversibly binds to activated caspases. Because the 660-VAD-FMK FLICA reagent becomes covalently coupled to the active enzymes, it is retained within the cell, while any unbound 660-VAD-FMK FLICA reagent diffuses out of the cell and is washed away. The remaining far red fluorescent signal is a direct measure of the active caspase enzyme activity present in the cell at the time the reagent was added. Cells that contain the bound FLICA can be analyzed by fluorescence microscopy or flow cytometry. Cells labeled with the FLICA reagent may be read immediately or preserved for 16 hours using the fixative. Unfixed samples may also be analyzed with Hoechst stain to detect changes in nuclear morphology.
Storage and Stability:Store powder at 4°C liquid at -20°C. Store other components at 4°C. Stable for 6 months For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.
