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rTEV蛋白酶(重组型) TEV Protease 烟草蚀纹病毒蛋白酶 1000U 800.00元特价

发表时间:2010-08-31  |  点击率:4639

 

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              本公司出口产品 TEV Protease 为国内科研客户提供物美价廉的rTEV蛋白酶 1000U  特价 800.00
TEV Protease烟草蚀纹病毒蛋白酶 
TEV Protease(TEV蛋白酶是来源于烟草蚀纹病毒(TEV)的Nla蛋白酶经改进后的50kDa的蛋白酶,经过设计后与天然TEV蛋白酶相比其稳定性更好。此蛋白酶被用来切除纯化后融合蛋白的亲和标签。TEV蛋白酶具有很强的位点特异性,能够识别EXXYXQ(G/S)的七氨基酸序列,普通的是ENLYFQG,其切割位点在谷氨酰胺和甘氨酸或丝氨酸之间。蛋白酶在G/S(或P1`)位点切割多种氨基酸序列,为切割后C端融合部分提供一个想要的N端氨基酸。在PH 7.0,30℃时可达到佳活性,但在PH 5.5-8.5和4-30的广泛范围内TEV蛋白酶皆有活性,使得反应条件的选择可根据目的蛋白的情况而修改。切割后也很容易利用其N端的HQ标签进行TEV蛋白酶清除。也可以从固定在树脂上的融合蛋白切除掉目的蛋白。
rTEV蛋白酶(重组型)
rTEV蛋白酶(重组型)是经过基因工程改造后的重组蛋白酶,该酶特异性识Glu-Asn-Leu-Tyr-Phe- Gln-Gly七氨基酸序列。rTEV蛋白酶与肠激酶U(EK)、Thrombin、FactorXa、SUMO等蛋白酶相比,具有高活性、高特异性的双重特点,rTEV蛋白酶因具高剪切活性和特异性,已成为融合蛋白表达后去除融合tag的蛋白酶。该酶经6×His标签纯化而得(含组胺酸标签),纯度达99%,剪切反应完毕后可通过His标签纯化树脂Ni-NTA Resin(Cat.No. P2010)去除。该酶在2-8-30温度、pH范围(6.0-8.5)反应条件下均具有活性(见下表)
               不同温度下剪切活性(%)
             4    16          21           30
 0.5 h              34     58           56                70
 1 h              58     80           78                90
 2 h                  71     99           99                99
 1 h              84     99           99                99
组分与保存条件:
货号     组分名称                     规格        数量               保存
P2060    rTEV 蛋白酶 (冻干粉)        1000U        1           -70/-20
P2060-A rTEV Buffer A                 1 ml        1                -20
P2060-B 500 X rTEV Buffer B            1ml         1                -20
备注:rTEV 蛋白酶保存条件:长期储存-70℃,可储存1年,-20℃,可储存6个月。 
 
剪切活性(%)电泳图谱
使用方法说明:
 1. 推荐使用溶液:50 mM NaH2PO4, 500mM NaCl, 1mM EDTA, 1mMDTT, 10%甘油,pH 8.0 buffer中进行。
   2. rTEV 蛋白酶 (冻干粉)1000U/支用1ml rTEV Buffer A溶解。
   3. rTEV与需要剪切的蛋白比例:1:100
   4. 推荐剪切时间仅供参考:用户可以根据自己研究的蛋白进行摸索,
 
剪切方法:
融合蛋白           1000 μg/ml
rTEVBuffe B         2 μl
rTEV蛋白酶        1 0μ
 

1000u       800.00

 

 

 

别名:TEV protease

表达方式: A DNA sequence encoding the TEV Protease (NP_062908) (Gly 2038- Gly 2280) was fused with polyhistidine tag at the N-terminus, with a Ser 2256 Asn mutation

种属:Human

表达宿主: E.Coli
纯度:> 95%, as determined by SDS-PAGE

内毒素:< 1.0 EU per 1μg cytokine as determined by the LAL method.

稳定性:Samples are stable for up to twelve months from date of receipt -70°C

预测N端: Met 1

分子量:The recombinant human TEV protease consists of 251 amino acids and has a predicted molecular mass of 28.6 kDa. As estimated in SDS-PAGE under reducing conditions.

缓冲液: Supplied as a 0.2μm filtered solution of 50mM Tris, 10%Sucrose,5mM DTT pH8.0

SDS-PAGE:
Usage Guide
储存方法:Store it under sterile conditions at -70℃. It is recommended that the protein be aliquoted for optimal storage and usage. Avoid repeated freeze-thaw cycles.

Reconstitution:Follow the instructions on the vial. Centrifuge the vial at 4℃ before opening to recover the entire contents.
Protein Description
TEV Protease is the 241 amino acid, 27 kDa catalytic domain of the nuclear inclusion a (NIa) protease from tobacco etch virus. As a 3C-type protease, this enzyme specifically recognizes and processes at the consensus sequence Glu-Asn-Leu-Tyr-Phe-Gln-Gly in cis and in trans, and cleavage occurs between the Gln and Gly residues. Unlike factor Xa, enteropeptidase or thrombin, TEV protease has not been found to cleave at unintended sites even when present at high concentration. Due to its high specificity and high activity rate within a wide range of pH and ionic strength conditions, TEV protease is an optimal and useful reagent for removing affinity tags from genetically engineered fusion proteins. However, a serious drawback of TEV protease is its autocatalytic activity at S2256 to generate a truncated enzyme with greatly diminished activity. Therefore, mutated TEV proteases have been produced for improved stability.

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