CleaTag™ TEV Protease

Catalog number :P5518

TEV Protease, also known as Tobacco Etch Virus (TEV) Protease, is a highly specific cysteine protease that recognizes the amino-acid sequence Glu-Asn-Leu-Tyr-Phe-Gln-(Gly/Ser) and cleaves between the Gln and Gly/Ser residues. It is often used for the removal of affinity purification tags such as maltose-binding protein (MBP) or poly-histidine from fusion proteins.

The enzyme is active over a wide range of pH 5.5-8.5 and 4-30 ℃, so that the choice of reaction conditions can be modified depending on the protein of interest.

TEV Protease has a 6xHis-tag for easy removal from a reaction using nickel affinity resins and has been engineered to improve thermal stability and decrease autolysis.

Overview

Description: CleaTag™ Recombinant TEV Protease(His-tagged)

Tested applications: Removal of affinity purification tags such as maltose-binding protein (MBP) or poly-histidine from fusion proteins

TEV protease encoded by the tobacco etch virus is a catalytic domain of the Nuclear Inclusion a (NIa) protein. It is consists of 241 amino acids with the molecular weight of 27kDa.

TEV recognizes the amino acid sequence of the general form E-X-X-Y-X-Q (or S)/X’, and cleaves between Q (or S)/X’. In this form X and X’ stand for any of the amino acid residues, except that X’ cannot be P. The optimal cleavage site is ENLYFQ/G. As having the absolute specificity and wildly using conditions like broad pH range and ionic strength, the TEV protease became more versatile than EK, thrombin and other protease used in biochemical applications, especially recombinant protein production. The optimal temperature for cleavage is 30°C; however, the enzyme can be used at temperatures as low as 4°C. Following digestion, TEV Protease can be removed from the reaction via the His tag sequence by Ni2+-chelate affinity chromatography.

Properties

Storage instruction: Long-term storage at -80 ℃, can be stored for 2 years; or a small amount of packaging stored at -20 ℃, can be stored for 6 months, avoid repeating freezing and thawing.

Database links: UniProt: P04517 (POLG_TEV) Activity domain: 2038 aa - 2273 aa

TEV protease encoded by the tobacco etch virus is a catalytic domain of the Nuclear Inclusion a (NIa) protein. It is consists of 241 amino acids(6 histidines and 235 aa enzyme) with the molecular weight of 27kDa.

TEV protease from Engibody Technology has the S219V mutation conferring improved activity, stability and resistance against self-cleavage (PMID 11809930).

HHHHHHHGESLFKGPRDYNPISSTICHLTNESDGHTT
SLYGIGFGPFIITNKHLFRRNNGTLLVQSLHGVFKV
KNTTTLQQHLIDGRDMIIIRMPKDFPPFPQKLKFREP
QREERICLVTTNFQTKSMSSMVSDTSCTFPSSDGIFWKHW
IQTKDGQCGSPLVSTRDGFIVGIHSASNFTNTNNYFTSV
PKNFMELLTNQEAQQWVSGWRLNADSVLWGGHKVFMVK
PEEPFQPVKEATQLMN

Applications

Application Images

Protocols: 1. Configurate the following reaction system in the EP tube:

Fusion protein:1mg

TEV Protease:10U

2. Incubate at suitable temperature after mixing the above reaction system. It is recommended that the reaction conditions of enzyme digestion is recommended at 4℃ overnight.

3. After the enzyme digestion, a small amount of samples can be selected for SDS-PAGE analysis.If you need to remove the TEV Protease in the reaction system after the enzyme digestion,using histidine tags purification resin affinity chromatography .

TEV Protease contains a 6x His-Tag for easy removal from a reaction using Ni agarose Resin (engibody,Cat no: P3403), or Ni-NTA Magnetic Beads (engibody,Cat no: P0030)

CleaTag™ SUMO Protease

Catalog number	P5519
Description	CleaTag™ SUMO Protease(Recombinant, his-tagged)
Application Images	Protein X was expressed as a SUMO fusion protein. To remove the SUMO tag, the SUMO-X fusion protein was incubated with SUMO protease at a ratio of 200:1 (w:w); after 10 minutes, >99% of the protein was cleaved.

Synonym	Yeast SUMO protease,Ubiquitin-like-specific protease 1,Small Ubiquitin-like Modifier Protease,Ulp1
Uniprot ID	Q02724 (ULP1_YEAST)
Origin of species	Yeast (Saccharomyces cerevisiae)
Source	Yeast SUMO protease variant corresponding to the active SUMO protease domain (amino acids Leu403-Lys621) expressed recombinantly in and purified from Yeast
Molecular Weight	28kDa
Storage	Upon receipt, store SUMO protease at -80℃ and buffers at -20℃. Aliquot after the first thawing. Please avoid repeated freeze/thaw cycles. Stable for 24 months when stored properly.
Fusion Tag(s)	N-terminal 2*polyhistidine tag
Purity	>95%
Buffer	12.5 mM Tris (pH 7.5), 250 mM NaCl, 125 mM Imidazole, 1 mM DTT, 50 % glycerol
Activity	Cutting will be dependent on spacing between the SUMO tag and the rest of the protein. Hence, it is hard to define activity. In most cases, 1 hour incubation at a ratio 1:200 at room temperature will result in >99% cleavage of the SUMO tag.
Shipped	Dry ice

Amino Acid Sequence

MGSSHHHHHHSSGGTENLYFQGHM

LVPELNEKDDDQVQKALASRENTQLM

NRDNIEITVRDFKTLAPRRWLNDTIIEFF

MKYIEKSTPNTVAFNSFFYTNLSERGYQ

GVRRWMKRKKTQIDKLDKIFTPINLNQ

SHWALGIIDLKKKTIGYVDSLSNGPNAM

SFAILTDLQKYVMEESKHTIGEDFDLIHLD

CPQQPNGYDCGIYVCMNTLYGSADAPL

DFDYKDAIRMRRFIAHLILTDALK

CleaTag™ Bovine Enterokinase

Catalog number	P4159
Description	CleaTag™ Recombinant Bovine Enterokinase (EK,his-tagged)
Tested applications	Fusion tag removal from the recombinant peptide fragment or protein that is preceded by the characteristic sequence DDDDK Cleavage or processing of engineered proteins that contain enzyme recognition sequence. Processing enzymes and proteins that are unstable in room temperature Enterokinase will not work if the recognition site is followed by a proline. rbovine EK with 6 × His-tag binds with Ni2+ affinity resin and was designed for removing from digestion system.
Highlights	High activity One unit is able to cleave 0.5 mg of a DDDDK-containing protein to 95% completion in 12 to 16 hours at 25°C. It is supplied as concentrated solution of > 5U/µL. High purity There are no any other protease contaminants nor animal-derived impurities. High specificity 1. Protease that cleaves specifically after a lysine preceded by four aspartic acids: Asp-Asp-Asp-Asp-Lys (DDDDK) 2. No detectable off-target cleavage when used at 2 - 25°C. Long-lasting superior performance. Retained enzymatic activity at 2 - 8°C. It allows low-temperature digestion to ensure stability of temperature-sensitive target proteins Compatible with the following reagents: Imidazole (<100 mM), NaCl (<50 mM), and glycerin (<5%).

CleaTag™ PreScission Protease

Catalog number	P4157
Description	CleaTag™ Recombinant PreScission Protease (GST-tagged)
Application Images
Specificity	PreScission Protease is a fusion protein of human rhinovirus (HRV) 3C protease and GST. HRV 3C Protease is encoded by human rhinovirus 14. It allows for on-column cleavage of GST tags and protein purification in one step. Specific cleavage – between the Gln and Gly residues of the recognition sequence LeuGluValLeuPheGln/GlyPro. Vectors encoding the HRV 3C protease recognition sequence pGEX-6P vectors pGEX-6P-1, pGEX-6P-2, and pGEX-6P-3 or other vectors encode the recognition sequence for this 3C protease.