Recombinant Tev Protease (His-tag) for Protein Purification
China Recombinant Tev Protease (His-tag) for Protein Purification, Find details about China Protein Purification Reagent, Protease His-Tag from Recombinant Tev Protease (His-tag) for Protein Purification
Basic Info.
Model NO.
G3409-5000U
Habit Appellation
Fine Chemicals
Application
Industry, Scientific Research
Property
Enzyme & Coenzyme
Trademark
Servicebio
Specification
5000U
Origin
China
Model NO.
G3409-5000U
Habit Appellation
Fine Chemicals
Application
Industry, Scientific Research
Property
Enzyme & Coenzyme
Trademark
Servicebio
Specification
5000U
Origin
China
Product Desciption
Recombinant TEV Protease (His-tag)
Product information
Product introduction
Recombinant TEV Protease is a protein expressing the cysteine Protease of Tobacco Etch Virus (TEV) with His tag in Escherichia coli. TEV Protease is a commonly used cysteine Protease, which can specifically recognize the heptopeptide sequence Glu-Asn-Leu-Tyr-Phe-Gln-Gly/Ser(E-N-L-Y-F-Q-G/S), and perform enzyme digestion between Gln(Q) and Gly/Ser(G/S) amino acid residues. The optimal digestion temperature of Recombinant TEV Protease is 30ºC, and the enzymatic activity of Recombinant TEV Protease is high in the range of 29-34ºC. However, when the temperature reaches or exceeds 37ºC, the enzymatic activity of Recombinant TEV Protease decreases sharply. In the actual operation, in order to preserve the structure and biological activity of the target protein as much as possible, it is recommended to use TEV Protease enzyme digestion overnight at 4ºC. The Recombinant TEV Protease is active in the pH range of 6.0 to 9.0, but loses its enzyme activity when the pH is less than or equal to 5.TEV Protease still has high activity in 400mM imidazole. Therefore, for the purification of target protein containing His label by nickel column, TEV Protease can be directly added into the purified target protein solution containing high concentration of imidazole. At 4ºC, the imidazole is removed by dialysis and the enzyme digestion is performed at the same time. It is also possible to remove HIS label by enzymatic digestion with TEV Protease after dialysis. The TEV Protease with His tag in the target protein solution after enzyme digestion and the excised His tag can be removed by binding with nickel column.
Application: It is often used to remove MBP, GST, HIS or other tagged proteins from fusion proteins. TEV Protease has its own label and can bind to the nickel column.
Source: Tobacco Etch Virus (TEV) cysteine protease, recombinant expression of Escherichia coli, containing HIS label.
Enzyme activity definition: at 30ºC, pH8.0, 3μg of substrate (containing Recombinant Tev Protease cleavage site) can be cleaved within 1h more than 85% of the required enzyme activity unit.
Inactivation or inhibition: Common serine Protease inhibitors such as PMSF, AEBSF, Bestatin, Pepstatin and EDTA do not inhibit TEV Protease activity. Protease inhibitors targeting cysteine residues, such as NEM or IAA, can significantly inhibit the activity of TEV Protease. TEV Protease still maintain higher enzyme activity in higher concentration of imidazole.
Purity and concentration: SDS-PAGE test purity ≥95%, 10 U/μL.
Recombinant TEV protease storage solution:50 mM Tris-HCl,250 mM NaCl,1 mM EDTA,5 mM DTT,50% (v/v) Glycerol,pH 8.0.
10×TEV Buffer:500 mM Tris-HCl,500 mM NaCl,5 mM EDTA,10 mM DTT,pH 8.0.
Storage and transportation
Transprot with wet ice; store at -20ºC, valid for 12 months.
Specificaiton
Operating steps:
1.Portenase digestion system recommendation
2. The reaction system was placed at 30ºC for 1-4h. If the target protein is not stable at 30ºC, the reaction system can be placed at 4ºC overnight for enzymatic digestion (about 16h).
3. SDS-PAGE assay was performed on a 10μL sample of digestion product to evaluate the digestion dosage and reaction conditions
Notes:
1. Enzyme products should be placed in an ice box or ice bath, and stored at -20ºC immediately after use. Separate storage is recommended.
2. For your safety and health, please wear a lab coat and disposable gloves to operate.
Figure 1: Effects of A+B fusion protein containing enzymatic digestion site of Recombinant TEV Protease (digestion condition 30ºC, pH 8.0, Enzymatic digestion for 0min, 10min, 20min, 30min, 1h, 2h, respectively, corresponding Lane Lane: 1-6, the mass ratio of substrate to enzyme added in the digestion system was 20:1).
Product information
| Product name | Cat. No. | Specification |
| Recombinant TEV Protease (His-tag) | G3409-5000U | 5000U |
Product introduction
Recombinant TEV Protease is a protein expressing the cysteine Protease of Tobacco Etch Virus (TEV) with His tag in Escherichia coli. TEV Protease is a commonly used cysteine Protease, which can specifically recognize the heptopeptide sequence Glu-Asn-Leu-Tyr-Phe-Gln-Gly/Ser(E-N-L-Y-F-Q-G/S), and perform enzyme digestion between Gln(Q) and Gly/Ser(G/S) amino acid residues. The optimal digestion temperature of Recombinant TEV Protease is 30ºC, and the enzymatic activity of Recombinant TEV Protease is high in the range of 29-34ºC. However, when the temperature reaches or exceeds 37ºC, the enzymatic activity of Recombinant TEV Protease decreases sharply. In the actual operation, in order to preserve the structure and biological activity of the target protein as much as possible, it is recommended to use TEV Protease enzyme digestion overnight at 4ºC. The Recombinant TEV Protease is active in the pH range of 6.0 to 9.0, but loses its enzyme activity when the pH is less than or equal to 5.TEV Protease still has high activity in 400mM imidazole. Therefore, for the purification of target protein containing His label by nickel column, TEV Protease can be directly added into the purified target protein solution containing high concentration of imidazole. At 4ºC, the imidazole is removed by dialysis and the enzyme digestion is performed at the same time. It is also possible to remove HIS label by enzymatic digestion with TEV Protease after dialysis. The TEV Protease with His tag in the target protein solution after enzyme digestion and the excised His tag can be removed by binding with nickel column.
Application: It is often used to remove MBP, GST, HIS or other tagged proteins from fusion proteins. TEV Protease has its own label and can bind to the nickel column.
Source: Tobacco Etch Virus (TEV) cysteine protease, recombinant expression of Escherichia coli, containing HIS label.
Enzyme activity definition: at 30ºC, pH8.0, 3μg of substrate (containing Recombinant Tev Protease cleavage site) can be cleaved within 1h more than 85% of the required enzyme activity unit.
Inactivation or inhibition: Common serine Protease inhibitors such as PMSF, AEBSF, Bestatin, Pepstatin and EDTA do not inhibit TEV Protease activity. Protease inhibitors targeting cysteine residues, such as NEM or IAA, can significantly inhibit the activity of TEV Protease. TEV Protease still maintain higher enzyme activity in higher concentration of imidazole.
Purity and concentration: SDS-PAGE test purity ≥95%, 10 U/μL.
Recombinant TEV protease storage solution:50 mM Tris-HCl,250 mM NaCl,1 mM EDTA,5 mM DTT,50% (v/v) Glycerol,pH 8.0.
10×TEV Buffer:500 mM Tris-HCl,500 mM NaCl,5 mM EDTA,10 mM DTT,pH 8.0.
Storage and transportation
Transprot with wet ice; store at -20ºC, valid for 12 months.
Specificaiton
| Component Number | Component | G3409-5000U |
| G3409-1 | Recombinant TEV Protease (His-tag) (10 U/μL) | 500 μL |
| G3409-2 | 10×TEV Buffer | 3×1 mL |
| G3409-3 | Control Protein | 100 μL |
| Manual | 1 | |
Operating steps:
1.Portenase digestion system recommendation
| Component | Volume |
| 10×TEV Buffer | 5 μL |
| Interest protein | 30 μg |
| Recombinant TEV Protease (His-tag) (10 U/μL) | 1 μL |
| ddH2O | To 50 μL |
3. SDS-PAGE assay was performed on a 10μL sample of digestion product to evaluate the digestion dosage and reaction conditions
Notes:
1. Enzyme products should be placed in an ice box or ice bath, and stored at -20ºC immediately after use. Separate storage is recommended.
2. For your safety and health, please wear a lab coat and disposable gloves to operate.
Figure 1: Effects of A+B fusion protein containing enzymatic digestion site of Recombinant TEV Protease (digestion condition 30ºC, pH 8.0, Enzymatic digestion for 0min, 10min, 20min, 30min, 1h, 2h, respectively, corresponding Lane Lane: 1-6, the mass ratio of substrate to enzyme added in the digestion system was 20:1).
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