CleaTag™ TEV Protease

Overview

Description

CleaTag™ Recombinant TEV Protease(His-tagged)

Tested applications

Removal of affinity purification tags such as maltose-binding protein (MBP) or poly-histidine from fusion proteins

TEV protease encoded by the tobacco etch virus is a catalytic domain of the Nuclear Inclusion a (NIa) protein. It is consists of 241 amino acids with the molecular weight of 27kDa.

TEV recognizes the amino acid sequence of the general form E-X-X-Y-X-Q (or S)/X’, and cleaves between Q (or S)/X’. In this form X and X’ stand for any of the amino acid residues, except that X’ cannot be P. The optimal cleavage site is ENLYFQ/G. As having the absolute specificity and wildly using conditions like broad pH range and ionic strength, the TEV protease became more versatile than EK, thrombin and other protease used in biochemical applications, especially recombinant protein production. The optimal temperature for cleavage is 30°C; however, the enzyme can be used at temperatures as low as 4°C. Following digestion, TEV Protease can be removed from the reaction via the His tag sequence by Ni2+-chelate affinity chromatography.

Properties

Storage instruction

Long-term storage at -80 ℃, can be stored for 2 years; or a small amount of packaging stored at -20 ℃, can be stored for 6 months, avoid repeating freezing and thawing.

Database links

UniProt: P04517 (POLG_TEV)   Activity domain: 2038 aa - 2273 aa

TEV protease encoded by the tobacco etch virus is a catalytic domain of the Nuclear Inclusion a (NIa) protein. It is consists of 241 amino acids(6 histidines and 235 aa enzyme) with the molecular weight of 27kDa.

TEV protease from Engibody Technology has the S219V mutation conferring improved activity, stability and resistance against self-cleavage (PMID 11809930). 

Applications

Application Image

Protocols

1. Configurate the following reaction system in the EP tube:

Fusion protein:1mg

TEV Protease:10U
 

2. Incubate at suitable temperature after mixing the above reaction system. It is recommended that the reaction conditions of enzyme digestion is recommended at 4℃ overnight.
 

3. After the enzyme digestion, a small amount of samples can be selected for SDS-PAGE analysis.If you need to remove the TEV Protease in the reaction system after the enzyme digestion,using histidine tags purification resin affinity chromatography .

TEV Protease contains a 6x His-Tag for easy removal from a reaction using Ni agarose Resin (engibody,Cat no: P3403), or Ni-NTA Magnetic Beads (engibody,Cat no: P0030)