Fluorescent protease assay, Fluoro™
Supplier: G Biosciences
The Fluoro™ Protease Assay Kit is designed for the quantitative determination of proteases present in a protein sample. The assay uses fluorescein isothiocyanate (FITC)-labelled casein as a general protease substrate. The fluorescein label on the FITC-casein is highly quenched. When the proteases present in the sample of interest digest the FITC-casein substrate into smaller peptides, the quenching of the fluorescence label is relieved and the fluorescence of the substrate is increased. The fluorescence of the FITC-labeled peptide is measured with excitation at 485 nm and emission at 530 nm to determine protease activity. The kit detects picogram level of proteases present in the sample.
The kit is supplied with chemically stabilised MSG-Trypsin™ for use as a general protease control; however, other specific protease standard controls can be used. MSG-Trypsin™ is an ultra-pure trypsin from porcine pancreas, modified by methylation followed by TPCK treatment and is extremely resistant to autolysis. The kit components are sufficient for 1000 assays in a microtitre plate format.
Resorufin-Casein Protease Substrate: A colorimetric substrate that when treated with proteases releases resorufin that has an absorbance of 570 nm (Supplied lyophilised). FITC-Casein Protease Substrate: A fluorescent (fluorescein isothiocyanate (FITC)) substrate that when treated with proteases releases FITC that has an excitation at 485 nm and emission at 535 nm (Supplied lyophilised).
Ordering information: Accessories must be ordered separately.
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