Testing Data
(Jurkat cells were divided into treated and untreated pools and stained with 660-YVAD-FMK. After spiking FLICA into the cell culture media, Jurkat cells were treated with a negative control (left) or Nigericin (20 uM, catalog #6698) to induce caspase-1 activity (right) for 24 hours. Cells were washed 3 times and read on an Accuri C6 flow cytometer. Treatment with the negative control induced caspase-1 activity in only 6.8% of the cell population (left), whereas treatment with Nigericin induced caspase-1 activity in 57.3% of the experimental cells (right). This is a ratio of 8:1.)
Background: Caspases play important roles in apoptosis and inflammation. The FLICA assay 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.
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Product Notes
The FLICA 660 Caspase-1 (Catalog #AAA258044) is an Assay Kit and is intended for research purposes only. The product is available for immediate purchase. It is sometimes possible for the material contained within the vial of "FLICA 660 Caspase-1, Assay Kit" to become dispersed throughout the inside of the vial, particularly around the seal of said vial, during shipment and storage. We always suggest centrifuging these vials to consolidate all of the liquid away from the lid and to the bottom of the vial prior to opening. Please be advised that certain products may require dry ice for shipping and that, if this is the case, an additional dry ice fee may also be required.Precautions
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