|Description||PreScission Protease is a fusion protein of glutathione S-transferase (GST) and human rhinovirus (HRV) type 14 3C protease . The protease specifically recognizes a subset of sequences which include the core amino acid sequence Leu-Phe-Gln/Gly-Pro cleaving between the Gln and Gly residues . Substrate recognition and cleavage are likely to be dependent not only upon primary structural signals, but also upon the secondary and tertiary structures of the fusion protein as well.|
|Biological Activity||Unit Definition:
One unit is defined as the amount of enzyme needed to cleave 100 μg of fusion protein in 16 hours to 90% completion at 5°C in a buffer containing 50 mM Tris-HCl, pH 7.0, 150 mM NaCl, 1 mM EDTA, and 1 mM DTT.
50 mM Tris-HCl, pH 7.0 (at 25 °C), 150 mM NaCl, 1 mM EDTA, 1 mM dithiothreitol. Chill to 5 °C prior to use.
|Reaction Buffer||Cleavage Buffer:
50mM Tris-HCl, pH7.0 (at 25°C), 150 mM NaCl, 1 mM EDTA, 1 mM dithiothreitol. Chill to 5°C prior to use.
|Physical Appearance||Sterile colorless liquid.|
|Usage||This material is for research, laboratory or further evaluation purposes. NOT FOR HUMAN USE|
|Storage||Should be stored in small aliquots at -20°C to -80°C for long term.|
|Note||This material is offered for research, laboratory or further evaluation purposes. NOT FOR HUMAN USE.|
|During cleavage reactions, it is recommended that samples be removed at various time points and analyzed by SDS-PAGE to estimate the yield, purity, and extent of digestion. The amount of PreScission Protease, temperature and length of incubation required for complete digestion of a given GST fusion partner may vary depending on the fusion partner. Optimal conditions for each fusion should be determined in pilot experiments. Digestion may be improved by adding TritonTM X-100, TweenTM 20, NonidetTM, or NP40 to a concentration of 0.01%. Concentrations of these detergents up to 1% do not inhibit PreScission Protease.
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