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RM5-0021 EN-TEV Protease $160

EN-TEV Protease

Product Name: EN-TEV Protease
Catalog #:  RM5-0021
Manufacture:  LD Biopharma, Inc.
Intruduction: TEV protease recognizes a linear epitope of the general form E-Xaa-Xaa-Y -Xaa-Q-(G/S), with cleavage occurring between Q and G or Q and S. The most commonly used sequence is ENLYFQ / G. A more stable mutant (S219N) of TEV protease (EN-TEV) was expressed and purified from E.coli with an N-terminal poly histidine tag was described by the Doudna laboratory [Lucast et al., 2001]. The protease is used to cleave affinity tag from fusion protein. The optimal temperature for cleavage is 30 °C. EN-TEV could be easily removed from the cleavage reaction by affinity chromatography using the polyhistidine tag at N-terminal of the EN-TEV enzyme.
Gene Symbol:  TEV 
Accession Number:  ABF71454.1
Species:  Tobacco Etch Virus (TEV)
Package Size:  50 µg / Vial   
Composition: 1.0 mg/ml, sterile-filtered, in 50 mM pH 8.0 Tris-HCl Buffer, with 0.5mM EDTA and1mM DTT.
Storage: In Liquid. Keep at -20°C for long term storage. Product is stable at 4 °C for 2-3 weeks.
Key Reference: Kapust, R. B., Tözsér, J., Fox, J. D., Anderson, D. E., Cherry, S., Copeland, T. D., and Waugh, D. S. Tobacco etch virus protease: Mechanism of autolysis and rational design of stable mutants with wild-type catalytic proficiency. Prot. Eng. 14: 993-1000 (2001).

The "standard" reaction buffer for TEV protease is 50 mM Tris-HCl (pH 8.0), 0.5 mM EDTA and 1mM DTT. The duration of the cleavage reaction is typically overnight, although lots of cleavage will happen in the first few hours and prolonged incubation times may not lead to proportional increases in cleavage. TEV protease is maximally active at 34 °C, but we recommend performing the digest at room temperature (20 °C) or 4 °C. TEV protease is only three-fold less active at 4 °C than at 20 °C (Nallamsetty et al., manuscript in preparation).

Typically, a good rule of thumb for initial test of digestion is 1 ug TEV enzyme per 50 - 100ug target protein ratio. Perform a small-scale reaction first, if possible, to gauge the efficiency of processing.

Quality Control:

1. Purity: > 98% by SDS-PAGE.

2. Activity: Completely digestion 50ug of T7 Tag-HBxAg protein using 1ug TEV enzyme ( in 1: 50 ratio) in either 3 hours at 37 °C or 12 hours at 4 °C.

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