TEV Protease, 2000U

Activity and Specificity

BP^TMTEV Protease has a specific activity of at least 10,000 units/mg, using the conventionally defined activity unit (One unit cleaves >85% of 3 μg control substrate in 1 h at 30^oC). In practice, 10,000 units (1mg) of BP^TMTEV Protease cleaves >90% of 100 mg of a control target protein at 4^oC in 16 hours. No non-specific cleavage has been observed under the same condition when BP^TMTEV Protease and the control target protein was mixed at 1:10 ratio.

Component and Formulation

BP^TMTEV Protease: 2 0,000U/ml in 25 mM Tris-HCl, pH 8.0, 50 mM NaCl, 1 mM TCEP, and 50% glycerol.

Cleavage Condition

It is recommended to test BP^TMTEV Protease cleavage with a protease-to-target protein ratio of 1:100 (w/w) or 1 unit of BP^TMTEV to 10 mg of target protein in a buffer suitable for the target protein at 4^oC overnight, with the target protein concentration at 1-2 mg/ml. In most cases, >90% of target protein is cleaved with a T BP^TMTEV -to-target protein ratio of 1:50 to 1:200 or 1 unit BP^TMTEV to 5-20 mg of target protein (as shown in Figure 1). The efficiency of cleavage may vary due to the sequences around the cleavage site, the conformation and the solubility of the target protein. Due to its high specificity, more BP^TMTEV Protease (at 1:10 ratio) or longer cleavage time (over a weekend) at higher temperature (37^oC) can be used to achieve high cleavage efficiency without non-specific cleavage of target proteins.

Removal of BP^TMTEV Protease after Cleavage

TEV Protease contains both GST and His tags. After cleavage of the target protein, BP^TMTEV Protease is easily removed along with the tags from the cleavage reaction by affinity chromatography using Ni-chelating resin for His-tagged target protein or GSH resin for GST-tagged target protein.