Less than 0.1EU/μg of rPSP as determined by LAL method.
Reconstitution
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 or NonidetTM P40 to a concentration of 0.01%. Concentrations of these detergents up to 1% do not inhibit PreScission Protease.
Category
Enzymes
Background
PreScission protease is a cysteine protease derived from human rhinovirus-HRV3C Protease. rPP is a fusion protein of glutathione S-transferase (GST) and human rhinovirus (HRV) type 14 3C protease. It specifically recognizes the amino acid sequences which include the core site of Leu-Phe-Gln-Gly-Pro and cleaves 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 super structures of the fusion protein. rPP works most effective at 4℃ and can digest substrates at room temperature as well.