protein

EZCut TEV Protease, Recombinant

MBS844472

0.1 mg

150 USD

Recombinant Protein

EZCut TEV Protease, Recombinant

[EZCut TEV Protease]

[Nuclear inclusion protein A; NIa protein]

The EZCut TEV Protease has an activity of >=10,000 units/mg.

>=95% by SDS-PAGE

Liquid. 1mg/ml solution in 0.1 M Tris-HCl, 0.5 M NaCl, 20% glycerol, 5 mM DTT and 0.5 mM EDTA, pH 8.0

1 mg/ml

Store at -80°C. Stable for at least 1 year as supplied. It may be further diluted to 0.1-0.5 mg/ml with 0.1M Tris-HCl, 0.5M NaCl, 20% glycerol, and stored at -80°C in aliquots. Avoid repeated freezing and thawing cycles.

SDS-PAGE; HPLC

Recombinant EZCu TM TEV Protease can be used to cleave solubility, secretion, detection, and purification affinity tags from recombinant proteins containing TEV protease-specific cleavage site resulting in the target protein with only one extra amino acid at N-terminus in comparison to other endopeptidases. The extra amino acid (Glycine/Serine) is a smaller amino acid and may not cause significant changes to protein structure.

SDS-PAGE

Testing Data

Unit Definition: One unit is defined as the amount of EZCut TEV Protease required to cleave >90% of 10 ug a control protein substrate (containing an ENLYFQS sequence at N-terminus) in 1 h at 37°C.

Source: E.coli. Cleavage Protocol: In order to find the optimum cleavage conditions for a target fusion protein, it is recommended to run preliminary cleavage reactions at a small scale. Successful cleavage with TEV protease is dependent upon proper folding of the fusion protein that enables access of the TEV recognition sequence by the enzyme. The target fusion protein (1 mg/ml) containing TEV protease cleavage site should be purified to homogeneity. Recommended cleavage buffer is 50 mM Tris, buffer, 0.1 M NaCl, pH 8.0. 1.To 10 ul of target protein in an Eppendorf tube add 0.05-0.5 ug (0.5-5 U) of TEV protease. 2.Add 10 ul of freshly prepared 50 mM Tris, buffer, 0.1 M NaCl, pH 8.0 containing 5 mM DTT. Mix well by pipetting up and down; incubate at 37°C for 1 h. Analyze by SDS-PAGE. 3.Once optimum cleavage conditions are obtained, the reaction can be scaled up to cleave the entire amount of the target fusion protein. After completion of cleavage reaction, TEV can be removed by passing the entire mixture through Ni-chelating resin. Collect the flow through and wash with the resin with 50 mM Tris, buffer, 0.1 M NaCl, pH 8.0. Combine the washes with the flow through to obtain cleaved protein product. 4. Optimum temperature for TEV protease cleavage is 34°C. However, the reaction can also be carried out at 37°C or 30°C also. If the target protein is not stable at these temperatures, the cleavage reaction can also be performed at room temperature or 4°C. In such cases, the amount of TEV protease should be optimized for different target proteins

Molecular Biology Tools; Molecular Biology Proteins/EnzymesProteins and Enzymes; ProteasesProteins and Enzymes; Animal-Free Origin (AOF) Recombinant Proteins & EnzymesProteins and Enzymes; Proteins and Enzymes (A-Z)

Background: EZCut TEV Protease is a cysteine protease that recognizes the cleavage site of Glu-Xaa- Xaa-Y- Xaa-Gln-(Gly/Ser) and cleaves between Gln and Gly/Ser. The optimal sequence is Glu-Asn-Leu-Tyr-Phe-Gln-Ser/Glycine (ENLYFQS/G). It contains an enhanced form of a catalytic fragment of the NIa protein of Tobacco etch virus (TEV). TEV Protease is a restriction grade protease that has robust activity at 4°C with high specificity and great stability. The optimal temperature for cleavage with this enzyme is 34°C. The protease can be used for the removal of affinity tags from fusion proteins. It contains a C-terminal His tag and can be easily removed after cleavage reactions by passing the reaction through a Ni-chelating resin. EZCut TEV Protease is an improved version of TEV protease that is highly site-specific, highly active, and significantly more stable than native TEV protease, resulting in enhanced long-term activity.

28.6 kDa (2038-2279 aa + C-terminal poly-his tag).

0.1 mg

150 USD

1 mg

415