Recombinant PreScission Protease, Low Endotoxin (rPPase,LowEndotoxin)
3C protease, Picornain 3C, PSP
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.
Sterile colorless liquid.
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.
Less than 0.1 EU/ug of rPPase as determined by LAL method.
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.
Use a manual defrost freezer and avoid repeated freeze-thaw cycles.- 6 months from date of receipt, -20 to -70 °C as supplied.- 3 months, -20 to -70 °C under sterile conditions after opening.
One unit is defined as the amount of enzyme needed to cleave 100 ug 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.
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