Description
Product description
Recombinant TEV protease (rTEV Protease) is a recombinant protease modified and purified by genetic engineering, which not only maintains the functional activity of natural TEV Protease, but also shows stronger stability and specificity over a broad temperature range. rTEV Protease is a commonly tool enzyme for cutting affinity labels on fusion protein. It is specific to the seven amino acid sequence EXXYXQ↓(G/S), and the cutting site is between glutamine and glycine/serine. The commonly used seven amino acid sequence is Glu-Asn-Leu-Tyr-Phe-Gln↓-Gly. rTEV Protease is optimally active at pH 7.0, 30°C, but is active over a wide range of pH 6.0-8.5 and temperatures 4-30°C (see Table 1), allowing the choice of reaction conditions to be flexible depending on the target protein. In addition, the 6×His label of N-terminal can also be used to remove rTEV Protease by Ni-NTA resin to purify the target protein.
The product has been optimized on the basis of 20403ES, and compared with 20403ES, it has higher purity, stronger activity and lower E.coli gDNA residue, which is suitable for application with higher requirements for enzyme purity, activity and residue.
Specifications
|
Cat.No. |
|
|
Size |
1 KU / 10 KU / 50 KU |
|
Enzyme activity |
5 U/μL |
|
Unit Definition |
The amount of enzyme required to cut >85% of the 3 μg substrate in a 1×rTEV Buffer (50 mM Tris, pH 8.0, 0.1 mM EDTA, 1 mM DTT) at 30℃ for 1 h was defined as a unit of activity |
|
Protein purity |
≥95% |
|
Source |
Expressed in E. coli. |
Components
|
Components No. |
Name |
20427ES80 |
20427ES10 |
20427ES50 |
|
20427-A |
200 μL |
2×1 mL |
10 mL |
|
|
20427-B |
rTEV Buffer (10×) |
600 μL |
6 mL |
30 mL |
Storage
rTEV Protease, stored at -25~-15℃ for 6 months; Store at -85~-65℃ for 1 year.
rTEV Buffer, stored at -25~-15℃ for 1 year.
Note: It is recommended to store each component separately when receiving the goods or using it for the first time to avoid repeated freezing and thawing.
Instructions
1. The following reaction system is formulated in the EP tube:
|
Components |
Volume (μL) |
|
Fusion protein |
40 μg |
|
rTEV Protease (5 U/μL) |
4 μL |
|
rTEV buffer (10×) |
10 μL |
2. Incubation at 30℃ for 2 h or 4℃ overnight.
Table 1. rTEV Protease digestion activity at different temperatures
|
Incubation Time（h） |
Digestion activity at different temperatures (%) |
|||
|
4℃ |
16℃ |
21℃ |
30℃ |
|
|
1 |
34 |
58 |
56 |
85 |
|
2 |
58 |
80 |
78 |
90 |
|
3 |
71 |
99 |
99 |
99 |
|
3.5 |
84 |
99 |
99 |
99 |
Notes
1. Factors affecting natural TEV Protease activity:
1) PMSF and AEBSF (1 mM), TLCK (1 mM), Bestatin (1 mg/mL), EDTA (1 mM), Pestatin A (1 mM) and E-64 (3 mg/mL), while 20427ES was compatible with the above inhibitors;
2) Zn2+ ion concentration (>5 mM) also inhibited 20427ES activity;
3) Reagents that react with cysteine are potential inhibitors of 20427ES.
2. Please wear the necessary PPE, such lab coat and gloves, to ensure your health and safety.
3. This product is for research use only.
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