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Home Protein Tools Products TEV Protease

TEV Protease

The TEV protease recognition sequence with the highest catalytic efficiency is ENLYFQ ▼S; however, the amino acid in the P1’ position can also be G, A, M, C, or H (1).

Removal of affinity purification tags such as maltose-binding protein (MBP) or poly-histidine from fusion proteins
Optimal activity and stability for up to 24 months
Active in a wide range of buffers; optimal activity between pH 6.0 and 9.0.
High substrate specificity with no non-specific proteolysis
TEV Protease contains a 7x His-Tag for easy removal from a reaction using NEBExpress^® Ni Resin (NEB# S1428S), NEBExpress^® Ni Spin Columns (NEB# S1427S/L), or NEBExpress^® Ni-NTA Magnetic Beads (NEB# S1423S)

Catalog #	Concentration	Size	List Price	Your Price
P8112S	10,000 units/ml	1,000 units	$108.00	Sign In Sign In

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Product Information

TEV Protease is a highly specific cysteine protease. The TEV Protease recognition sequence with the highest catalytic efficiency is ENLYFQ ▼S; however, the amino acid in the P1’ position can also be G, A, M, C, or H (1). It is often used for the removal of affinity purification tags such as maltose-binding protein (MBP) or poly-histidine from fusion proteins. TEV Protease has a 7xHis-tag for easy removal from a reaction using nickel affinity resins and has been engineered to improve thermal stability and decrease autolysis.

Product Source

Cloned from Tobacco Etch Virus and expressed in E. coli.

Reagents Supplied

The following reagents are supplied with this product:

Show all

NEB # Component Name Component # Stored at (°C) Amount Concentration

P8112S -20

TEV Protease P8112SVIAL -20 1 x 0.1 ml 10,000 units/ml

TEV Protease Reaction Buffer B8035SVIAL -20 1 x 1 ml 10 X

This product is related to the following categories:: NEBExpress MBP Fusion and Purification System,; Bacterial E. coli Protein Expression,; Nickel Purification (His-tag),; Proteases Products,; Protein Purification,

This product can be used in the following applications:: Fusion Protein Cleavage,; Target Protein Insolubility ,; Protein Purification,; Protein Digestion,

Advantages and Features
Features
- Removal of affinity purification tags such as maltose-binding protein (MBP) or poly-histidine from fusion proteins
- Engineered to improve thermal stability and decrease autolysis
- High substrate specificity with no non-specific proteolysis
Properties & Usage

Unit Definition
1 unit of TEV Protease will cleave 2 µg of MBP-fusion protein, MBP5-TEV-paramyosin ΔSal, to 95% completion in a total reaction volume of 10 µl in 1 hour at 30°C in 50 mM Tris-HCl (pH 7.5 @ 25°C) with 0.5 mM EDTA and 1 mM DTT.
Reaction Conditions

1X TEV Protease Reaction Buffer
Incubate at 30°C

1X TEV Protease Reaction Buffer
50 mM Tris-HCl
0.5 mM EDTA
1 mM DTT
(pH 7.5 @ 25°C)

Storage Buffer

50 mM Tris-HCl
250 mM NaCl
1 mM TCEP
1 mM EDTA
50% Glycerol
pH 7.5 @ 25°C

Heat Inactivation
65°C for 10 min
Molecular Weight
Apparent: 28 kDa

Unit Assay Conditions
Two fold dilutions of TEV Protease are incubated with 2 μg MBP5-TEV-paramyosin ΔSal and 1X TEV Protease Reaction Buffer in a 10 µl reaction. The reaction mix is incubated at 30°C for 1 hour. Separation of reaction products are visualized by SDS-PAGE.
Product Notes
1. The TEV Protease recognition sequence with the highest catalytic efficiency is ENLYFQ ▼S; however, the amino acid in the P1’ position (S in the recognition sequence) can also be G, A, M, C, or H (1).
2. If the fusion protein sample contains > 2 M urea, > 0.5 M Guanidine hydrochloride, > 50 mM imidazole, pH values below 6 or above 9, or cysteine protease inhibitors then it will be necessary to dialyze the fusion protein before TEV Protease cleavage.
3. TEV protease can be used at high concentrations without the occurrence of non-specific proteolysis.
4. Reactions may be scaled-up linearly to accommodate larger reaction volumes.
References
1. Kapust, R.B. et al. (2002). Biochem. and Biophysical Research Comm.. 294, 949-955.

Protocols, Manuals & Usage
- Protocols
  Typical Reaction Conditions for TEV Protease (NEB #P8112)
  
  His-tag removal from protein using TEV Protease
Tools & Resources
- Selection Charts
  Protease Selection Chart
FAQs & Troubleshooting
- FAQs
  Can TEV Protease recognize and cleave a sequence other than Glu-Asn-Leu-Tyr-Phe-Gln-(Gly/Ser)?
  
  Is it necessary to dialyze the sample prior to cleavage with TEV Protease?
  
  Can TEV Protease be used at lower temperatures?
  
  Is TEV Protease compatible with protease inhibitors?
  
  Is TEV Protease compatible with different reaction buffers?
  
  Can TEV Protease be removed from the reaction after cleavage?
- Tech Tips
  The optimal pH range is 6.0 – 8.0
  
  Optimal activity achieved in ≤0.2M NaCl; however, the enzyme retains some activity in up to 2M NaCl
  
  Cleavage at lower temperatures, such as 4ºC, requires overnight incubation
  
  Inhibition occurs in the presence of ≥ 5 mM Zn²⁺, ≥ 1 mM Cu²⁺and ≥ 10 mM Co²⁺
  
  Compatible with 10mM MgSO₄, MnCl₂ and CaCl₂ and up to 100mM EDTA
  
  Compatible with the following protease inhibitors: aprotinin, benzamidine, leupeptin, pepstatin, PMSF
  
  Can be used at high concentrations with no non-specific proteolysis occurring
  
  Some substrates may require extended incubation (up to three days at either 4°C or 30°C)
  
  If cleavage is not complete, add more TEV protease after 24 hours and continue incubation
Citations & Technical Literature
Quality, Safety & Legal
- Quality Control Assays
  
  Quality Control tests are performed on each new lot of NEB product to meet the specifications designated for it. Specifications and individual lot data from the tests that are performed for this particular product can be found and downloaded on the Product Specification Sheet, Certificate of Analysis, data card or product manual. Further information regarding NEB product quality can be found here.
- Specifications & Change Notifications
  Specifications & Change Notifications
  The Specification sheet is a document that includes the storage temperature, shelf life and the specifications designated for the product. The following file naming structure is used to name these document files: [Product Number]_[Size]_[Version]
  
  P8112S_v1
- Certificate of Analysis
  The Certificate of Analysis (COA) is a signed document that includes the storage temperature, expiration date and quality controls for an individual lot. The following file naming structure is used to name these document files: [Product Number]_[Size]_[Version]_[Lot Number]
  
  P8112S_v1_10017701
  
  P8112S_v1_10028767
  
  P8112S_v1_10031124
  
  P8112S_v1_10035078
  
  P8112S_v1_10037481
  
  P8112S_v1_10039748
  
  P8112S_v1_10045949
  
  P8112S_v1_10048134
  
  P8112S_v1_10049722
  
  P8112S_v1_10052586
  
  P8112S_v1_10055989
  
  P8112S_v1_10061036
  
  P8112S_v1_10064916
  
  P8112S_v1_10067995
  
  P8112S_v1_10073045
  
  P8112S_v1_10077415
  
  P8112S_v1_10081140
  
  P8112S_v1_10088321
  
  P8112S_v1_10097171
  
  P8112S_v1_10102349
  
  P8112S_v1_10111204
  
  P8112S_v1_10115461
  
  P8112S_v1_10121429
  
  P8112S_v1_10132855
  
  P8112S_v1_10138065
  
  P8112S_v1_10142229
  
  P8112S_v1_10150502
- Safety Data Sheets
  The following is a list of Safety Data Sheet (SDS) that apply to this product to help you use it safely.
  
  TEV Protease
  
  TEV Protease Reaction Buffer
- Legal and Disclaimers
  
  This product is covered by one or more patents, trademarks and/or copyrights owned or controlled by New England Biolabs, Inc (NEB).
  
  While NEB develops and validates its products for various applications, the use of this product may require the buyer to obtain additional third party intellectual property rights for certain applications.
  
  For more information about commercial rights, please email us at busdev@neb.com.
  
  This product is intended for research purposes only. This product is not intended to be used for therapeutic or diagnostic purposes in humans or animals.
  
  New England Biolabs (NEB) is committed to practicing ethical science – we believe it is our job as researchers to ask the important questions that when answered will help preserve our quality of life and the world that we live in. However, this research should always be done in safe and ethical manner. Learn more.

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