Recombinant EN-TEV Protease Protein, ＞98% (SDS-PAGE)

Cat. No.: rp156465

GRADE & PURITY Carrier Free, Bioactive, ActiBioPure™, Azide Free, His Tag, ≥98%(SDS-PAGE), See COA

Expression System

E. coli

Accession #

P04517

Protein Tag

N-His

Expression system

E. coli

Endotoxin Concentration

<1.0 EU/μg

Bioactivity

Measured by its ability to cleave a fusion protein containing the recognition sequence Glu-Asn-Leu-Tyr-Phe-Gln-Gly/Ser , with the cleavage point after Gln. One unit of TEV protease cleaves > 85% of 3 μg of control substrate in 1 hour at pH 8.0 at 30°C.It is recommended that the cleavage for each fusion protein be optimized by varying the amount of Recombinant Viral TEV Protease, reaction time, or incubation temperature.Complete digestion of 50µg of fusion KGF-MBP tag protein using 1µg TEV enzyme (in 1/50 ratio) in either 1 hours at 30 °C or overnight at 4 °C. The "standard" reaction buffer for TEV protease is 50 mM Tris-HCl (pH 8.0), 0.5 mM EDTA and 1mM DTT.

Overview

TEV Protease is the 241 amino acid (aa), 27 kDa catalytic domain of the nuclear inclusion a (NIa) protein encoded by the potyvirus, tobacco etch virus (TEV). It may be used in biotechnology to cleave affinity tags from recombinant proteins, either co-translationally orin vitrofollowing purification. Its high specificity and activity at a wide range of pH and ionic strength make TEV Protease more versatile than many other proteases used for the same purpose. Unlike factor Xa, enteropeptidase or thrombin, TEV Protease has not been found to cleave at unintended sites, even when present at a high concentration. TEV Protease is a 3C-type protease that cleaves substrates with a consensus sequence of ENLYFQG. Cleavage occurs between Q and G. Since the final aa remains on the cleaved protein where it could potentially affect structure or function, substitution of a variety of aa have been tested. In order of efficiency, S, A, M, Y, D, N, E, K or L may be effectively used in place of G. Several of the remaining aa may also vary, giving a final consensus sequence of ExxYF(M)Q(E)/G(S, A or others) where aa in parenthesis are alternatives and x is any aa. The autocatalytic site of NIa at S2256 has been mutated to an N for improved stability of the protease.
Tobacco Etch Virus Protease is a highly site-specific cysteine protease that is found in the tags from fusion proteins. The optimal temperature for cleavage is 30°C. It is recommended that the cleavage for each fusion protein be optimized by varying the amount of recombinant viral TEV protease, reaction time, or incubation temperature. It can be removed by Ni2+ affinity resin.

Specifications

Product Name

Recombinant EN-TEV Protease Protein, ＞98% (SDS-PAGE)

Synonyms

NIa | P1 Protease | TEV Protease | Tobacco Etch Virus Protease

Grade

ActiBioPure™, Azide Free, Bioactive, Carrier Free, His Tag

Specifications & Purity

Carrier Free, Bioactive, ActiBioPure™, Azide Free, His-Tag, ≥98%(SDS-PAGE), See COA

Purity

＞98% (SDS-PAGE)

Bioactivity

Measured by its ability to cleave a fusion protein containing the recognition sequence Glu-Asn-Leu-Tyr-Phe-Gln-Gly/Ser , with the cleavage point after Gln. One unit of TEV protease cleaves > 85% of 3 μg of control substrate in 1 hour at pH 8.0 at 30°C.It is recommended that the cleavage for each fusion protein be optimized by varying the amount of Recombinant Viral TEV Protease, reaction time, or incubation temperature.Complete digestion of 50µg of fusion KGF-MBP tag protein using 1µg TEV enzyme (in 1/50 ratio) in either 1 hours at 30 °C or overnight at 4 °C. The "standard" reaction buffer for TEV protease is 50 mM Tris-HCl (pH 8.0), 0.5 mM EDTA and 1mM DTT.

Endotoxin Concentration

<1.0 EU/μg

Expression System

E. coli

Species

Tobacco etch virus (TEV)

Amino Acids

2038-2279 aa

Sequence

GHHHHHHHGESLFKGPRDYNPISSSICHLTNESDGHTTSLYGIGFGPFIITNKHLFRRNNGTLVVQSLHGVFKVKDTTTLQQHLVDERDMIIIRMPKDFPPFPQKLKFREPQREERICLVTTNFQTKSMSSMVSDTSSTFPSGDGIFWKHWIQTKDGQCGSPLVSTRDGFIVGIHSASNFANTNNYFTSVPKNFMELLTNQEAQQWVSGWRLNADSVLWGGHKVFMVKPEEPFQPVKEATQLMNRRRRR

Protein Tag

N-His

Accession #

P04517

Predicted molecular weight

28.6 kDa

SDS-PAGE

28.6 kDa

Molecule Type

Protein

Storage and Shipping

Shape

Liquid

Concentration

See COA

Storage

Store at -20°C,Avoid repeated freezing and thawing

Shipped In

Ice chest + Ice pads

Stability And Storage

Store at -20°C stable up to 1 year. Avoid freeze / thaw cycle.

Images

SDS-PAGE: Recombinant EN-TEV Protease protein (rp156465)

Recombinant EN-TEV Protease protein (rp156465) - SDS-PAGE
3 μg/lane of Recombinant EN-TEV Protease protein was resolved with SDS-PAGE under reducing (R) and non-reducing (N) conditions and visualized by Coomassie® Blue staining, showing a band at 28.6 kDa.