The FLICA 660 Caspase-1 BioAssay(TM) Kit is an in vitro assay that employs the fluorescent inhibitor probe 660-YVAD-FMK to label active caspase-1 enzyme 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(TM) kits are used by researchers seeking to quantitate apoptosis via caspase activity in cultured cells. The FLICA 660 Caspase-1 probe allows researchers to assess caspase-1 activation. The FLICA reagent 660-YVAD-FMK enters each cell and irreversibly binds to activated caspase-1. Because the 660-YVAD-FMK FLICA reagent becomes covalently coupled to the active enzyme, it is retained within the cell, while any unbound 660-YVAD-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-1 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. Excitation/Emission: 660nm/690nm Kit Components: *474021A: FLICA Caspase-1 Inhibitor (660-YVAD-FMK), 1 x1vial 474021B: 10X Cellular Wash Buffer, 1x15ml 474021C: Fixative, 1x6ml Storage and Stability: Store *474021A at 4C. After reconstitution store at -20C. Store other components at 4C. Stable for 6 months For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.
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