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Recombinant Tobacco Etch Virus Protease (rTEV)

Recombinant Tobacco Etch Virus Protease (rTEV)

货号: UA070003
价格: 320
规格: 1kU
介绍: -
其他: -
产品规格

  • 物种

    Tobacco Etch Virus

  • 分子别名

    rTEV,TEV,RTEV

  • 浓度

    1U/μ、10U/μl

  • 表达宿主

    E.coli

  • 分子量

    28 kDa (Reducing)

  • 纯度

    ＞98% by SDS-PAGE & RP-HPLC

  • 内毒素含量

    <1EU/μg

  • 标记

    Unconjugated

  • 标签

    His Tag

  • 性状

    Liquid

  • 缓冲体系

    20mM Tris-HCl, pH8.0, 150mM NaCl, 1mM EDTA, 5mM DTT, 50%(v/v) Glycerol

  • 储存条件

    · 12 months from date of receipt, -20 to -70 °C as supplied. 
    · 6 months, -20 to -70 °C under sterile conditions after reconstitution.
    · 1 week, 2 to 8 °C under sterile conditions after reconstitution.  
    · Please avoid repeated freeze-thaw cycles.

  • 稀释度

背景介绍

  • The tobacco etch virus (TEV) protease is a useful tool for the removal of fusion tags from recombinant proteins. TEV protease has a strict 7 amino acid cleavage recognition sequence of Glu-Asn-Leu-Tyr-Phe-Gln-Gly/Ser [ENLYFQ(G/S)] and cleavage occurs between the Gln and Gly/Ser residues, The most commonly used sequence is ENLYFQG. 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. Recombinant Tobacco Etch Virus Protease (rTEV) has (NIa) protease catalytic domain which corresponds to a molecular weight of 28 kDa. It is unique with high specificity and is active at low temperature. rTEV Protease has a 6*His-tag for easy removal from a reaction using nickel affinity resins and has been engineered to improve thermal stability and decrease autolysis.

产品组分
    1. TEV Protease;
    2. 10X TEV Protease Buffer + Salt: 500mM Tris-HCl, pH8.0, 500mM NaCl, 5mM EDTA, 10mM DTT;
    3. 10X TEV Protease Buffer – Salt: 500mM Tris-HCl, pH8.0, 5mM EDTA, 10mM DTT.
操作步骤

  • 1. Add the following to a microcentrifuge tube:Add the following to a microcentrifuge tube:

    Fusion Protein

    15μg

    10X rTEV Protease Buffer +/– Salt

    5μl

    rTEV Protease

    5U

    ddH2O

    To 50μl

    2. Mix and incubate at 30°C, Remove 10 μL aliquots at 1, 2, 4, and 6 hours.

    3. Analyze by SDS-PAGE. 


    Guidelines for Cleavage

    • For most fusion proteins, TEV Protease functions optimally in a reaction mixture without NaCl; however, conditions may be optimized by varying the NaCl concentration from 0 mM to 500 mM. Remember to take into account the contribution of salt from the enzyme and from your substrate. When setting up your cleavage reaction, use the appropriate 10X rTEV Protease Buffer +/- Salt.
    • Researchers need to optimize their specific reaction conditions. As an initial suggestion, 10 units of rTEV protease can be used per 30μg of target protein for 1 hour at 30 °C, or overnight at 2–8 °C. The cleavage efficiency can then be estimated by SDS-PAGE.
酶活定义
  • One unit of TEV protease cleaves ≥85% of 3 μg of control substrate in 1 hour at pH 8.0

  • 生物活性JSON

    • The substrate of 3 μg fusion protein was digested by enzyme, and the addition amount of rTEV Protease in the sample was 0.1U, 0.2U, 0.5U, 1U,1.5U and 2U, respectively. The substrate is a fusion protein with a molecular weight of 25 KD and reacted at 30 ℃ for 1 hour, and the target band of 23kD is produced after rTEV Protease digestion.

  • 电泳JSON

    • 1μg (R: reducing conditions, N: non-reducing conditions).

  • 反相高效液相色谱JSON(RP-HPLC)