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Caspase 1 Assay Kit (Fluorometric) (ab39412) provides a simple and convenient method for detecting the activity of caspase 1, which recognizes the sequence YVAD. The assay is based on detection of cleavage of substrate YVAD-AFC (AFC: 7-amino-4-trifluoromethyl coumarin). YVAD-AFC emits blue light (Em=400 nm); upon cleavage of the substrate by caspase-1 or related caspases, free AFC emits a yellow-green fluorescence (Em=505 nm), which can be quantified using a fluorometer or a fluorecence microtiter plate reader. Comparison of the fluorescence from a treated sample with an untreated control allows determination of the fold increase in caspase 1 activity.
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Caspase 1 (ICE, IL-1beta Converting Enzyme) is the prototypical member of the ICE family of proteases/caspases. Caspase 1 was first identified as a novel protease that generates the proinflammatory cytokine, interleukin 1 beta (IL-1 Beta), by cleaving the pro-interleukin after the Asp116 residue. In addition to its role in the activation of proinflammatory cytokines, Caspase 1 also appears to have functions in some, but not all, types of apoptosis in mammalian cells.
|2X Reaction Buffer||4 x 2ml|
|Cell Lysis Buffer||1 x 100ml|
|DTT||1 x 0.4ml|
|YVAD-AFC||1 x 0.5ml|
Titration of the caspase 1 (ab39901) (background signal subtracted, duplicates; +/- SD).
Badding M.A et al investigated the cytotoxicity of Indium-tin oxide (ITO). Previously,ITO had shown to be cytotoxic in cultured cells and pro-inflammatory in pulmonary animal models. ITO is used to make transparent conductive coatings for touch screens and liquid crystal display electronics. RAW cells were plated at 5 x 105 cells/well and treated with SITO, LPS, or Min-U-sil (cells were first primed with LPS). Cells were washed, lysed, and 100 μg of lysates were assayed using Caspase 1 assay kit (ab39412). PBS was used as a control. All conditions were run in duplicate wells and three independent experiments were performed for each time point.
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