Proteases are naturally present in all organisms. These enzymes are involved in a multitude of physiological reactions from simple digestion of food proteins to highly regulated cascades. Proteases can either break specific peptide bonds (limited proteolysis), depending on the aa sequence of a protein, or break down a complete peptide to amino acids (unlimited proteolysis). The activity can be a destructive change (abolishing a protein's function), an activation of a function (preform to mature form) or it can be a signal in a signaling pathway.Sample Type:Cell and tissue culture supernatants, urine, plasma and serum, as well as many other biological fluids, growth medium, food samples, etc.Intended Use:Protease Activity Assay Kit is designed for the quantitative determination of proteases present in the protein sample.Sensitivity:<500pg/wellTest Principle:The assay uses fluorescein isothiocyanate (FITC)-labeled casein as a general protease substrate. The fluorescein label on the FITC-Casein is highly quenched. Upon digestion by proteases present in the sample the FITC-Casein substrate is cleaved into smaller peptides which abolishes the quenching of the fluorescence label. The fluorescence of the FITC-labeled peptide fragments is measured at Ex/EmH5/530nm. The kit is supplied with Mass Spectrometry Grade (MSG), chemically stabilized Trypsin for use as a general protease control. However, other protease standard controls can also be used. This kit is easy to use and can detect ≤500pg/well Trypsin present in the sample.Kit Components:Protease Assay Buffer: 1x25mlProtease Substrate (lyophilized): 1x1vialFITC Standard (25uM): 1x200ulPositive Control (lyophilized): 1x1vialStorage and Stability:Store components at 4°C. Stable for 6 months For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.
