The MaxDiscovery™ Caspase-1 Detection Kit (Colormetric) is a colorimetric microplate-based assay to directly measure caspase-1 activation in cultured cells. This kit enables biomedical researchers to determine caspase-1 levels in cultured cells from a broad range of cell lines. The assay utilizes a simple, proven enzymatic assay to detect activated enzyme. The kit is designed to be used with a microplate reader and contains sufficient materials to test 96 samples. The MaxDiscovery™ Caspase-1 Assay Kit measures activated caspase-1 enzyme using a direct, plate-based, colorimetric reaction. The kit contains a substrate solution containing a caspase-specific tetrapeptide substrate, ac-Tyr-Val-Ala-Asp-pNA. The zymogen procaspase-1 has no reactivity towards this substrate. Active caspase-1 specifically cleaves the pNA group from the peptide moiety. The free pNA group has a yellow color, which strongly absorbs light at 405 nm; the color is also visible in the wells. Mammalian cells are growth in culture wells and treated. The treated cells are lysed in the wells (after removing the cell culture medium) using a cell lysis buffer provided with the kit. The lysate is then directly added to wells in a 96-well reaction microplate (provided) containing the ac-Asp-Glu-Val-Asp-pNA substrate solution. Caspase-1 activity is then measured using a plate reader for the increase in absorbance at 405 nm in 30 minutes.