Product Class: Other

TEV Protease
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Item# Description List Price Web Price Qty
P8112S TEV Protease - 1,000 units $142.00
$127.80
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*On-line ordering is for Canadian customers only. Web pricing is applicable only to orders placed online at www.neb.ca

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    Product Introduction

    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.

    • 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

    Product Information

    Description

    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. 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.
    Product Categories:
    Proteases Products
    Applications:
    Protein Expression Approaches ,
    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 Temperature

      -20°C

      Storage Conditions

      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 optimum sequence for the TEV protease cleavage site is Glu-Asn-Leu-Tyr-Phe-Gln-(Gly/Ser). However, it has been reported that other amino acid residues can be used in the P1´ position instead of Ser or Gly. These alternative P1´ amino acids include Ala, Met, and Cys (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

      1. Typical Reaction Conditions for TEV Protease (NEB #P8112)
      2. His-tag removal from protein using TEV Protease

      Tools & Resources

      Selection Charts

      • Protease Selection Chart

      Faqs & Troubleshooting

      FAQs

      1. Can TEV Protease recognize and cleave a sequence other than Glu-Asn-Leu-Tyr-Phe-Gln-(Gly/Ser)?
      2. Is it necessary to dialyze the sample prior to cleavage with TEV Protease?
      3. Can TEV Protease be used at lower temperatures?
      4. Is TEV Protease compatible with protease inhibitors?
      5. Is TEV Protease compatible with different reaction buffers?
      6. Can TEV Protease be removed from the reaction after cleavage?

      Troubleshooting

      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

      Quality, Safety & Legal

      Quality Assurance Statement

      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

      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]

      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]

      Safety DataSheets

      The following is a list of Safety Data Sheet (SDS) that apply to this product to help you use it safely.

      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 contact NEB's Global Business Development team at [email protected].

      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.

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