P8112S, TEV Protease - 1000 units

TEV Protease is a highly specific cysteine protease. The TEV Protease recognition sequence with the highest catalytic efficiency is ENLYFQ ▼S; however, the amino acid in the P1’ position can also be G, A, M, C, or H. It is often used for the removal of affinity purification tags such as maltose-binding protein (MBP) or poly-histidine from fusion proteins. TEV Protease has a 7xHis-tag for easy removal from a reaction using nickel affinity resins and has been engineered to improve thermal stability and decrease autolysis. 

Product Source

Cloned from Tobacco Etch Virus and expressed in E. coli.

Advantages and Features

Features

• Removal of affinity purification tags such as maltose-binding protein (MBP) or poly-histidine from fusion proteins
• Engineered to improve thermal stability and decrease autolysis
• High substrate specificity with no non-specific proteolysis 

• Properties & Usage

Unit Definition

1 unit of TEV Protease will cleave 2 µg of MBP-fusion protein, MBP5-TEV-paramyosin ΔSal, to 95% completion in a total reaction volume of 10 µl in 1 hour at 30°C in 50 mM Tris-HCl (pH 7.5 @ 25°C) with 0.5 mM EDTA and 1 mM DTT.

Reaction Conditions

1X TEV Protease Reaction Buffer
Incubate at 30°C

1X TEV Protease Reaction Buffer
50 mM Tris-HCl
0.5 mM EDTA
1 mM DTT
(pH 7.5 @ 25°C)

Storage Buffer

50 mM Tris-HCl
250 mM NaCl
1 mM TCEP
1 mM EDTA
50% Glycerol
pH 7.5 @ 25°C

Heat Inactivation

65°C for 10 minutes

Molecular Weight

Apparent: 28 kDa

Unit Assay Conditions

Two fold dilutions of TEV Protease are incubated with 2 μg MBP5-TEV-paramyosin ΔSal and 1X TEV Protease Reaction Buffer in a 10 µl reaction. The reaction mix is incubated at 30°C for 1 hour. Separation of reaction products are visualized by SDS-PAGE.