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G3409-5000U Recombinant TEV Protease His-tag

Specification: 5000ul
Usage: Remove MBP, GST, HIS or other tagged proteins from fusion proteins.
Storage and Transportation: Transport with wet ice; store at -20, valid for 12 months.
u:
  • G3409-5000U
  • Servicebio

Product Description

Overview

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 30℃, and the enzymatic activity of Recombinant TEV Protease is high in the range of 29-34℃. However, when the temperature reaches or exceeds 37℃, 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 4℃. 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 4℃, 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 30℃, 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.


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

Product Usage


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

2. The reaction system was placed at 30℃ for 1-4h. If the target protein is not stable at 30℃, the reaction system can be placed at 4℃ 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 -20℃ immediately after use. Separate storage is recommended.

2. For your safety and health, please wear a lab coat and disposable gloves to operate.


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