rTEV蛋白酶(重組型) TEV Protease 煙草蝕紋病毒蛋白酶 1000U 800.00元*
更新時間:2010-08-31 | 點(diǎn)擊率:6080
上海索寶生物科技有限公司
地址:上海市徐匯區(qū)欽江路15號14層
:200233
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工業(yè)客戶(大批量生產(chǎn))請: 馬玉濤
本公司出口產(chǎn)品 TEV Protease 為國內(nèi)科研客戶提供物美價廉的rTEV蛋白酶 1000U * 800.00
TEV Protease(TEV蛋白酶)是來源于煙草蝕紋病毒(TEV)的Nla蛋白酶經(jīng)改進(jìn)后的50kDa的蛋白酶,經(jīng)過設(shè)計后與天然TEV蛋白酶相比其穩(wěn)定性更好。此蛋白酶被用來切除純化后融合蛋白的親和標(biāo)簽。TEV蛋白酶具有很強(qiáng)的位點(diǎn)特異性,能夠識別EXXYXQ(G/S)的七氨基酸序列,普通的是ENLYFQG,其切割位點(diǎn)在谷氨酰胺和甘氨酸或絲氨酸之間。蛋白酶在G/S(或P1`)位點(diǎn)切割多種氨基酸序列,為切割后C端融合部分提供一個想要的N端氨基酸。在PH 7.0,30℃時可達(dá)到佳活性,但在PH 5.5-8.5和4-30℃的廣泛范圍內(nèi)TEV蛋白酶皆有活性,使得反應(yīng)條件的選擇可根據(jù)目的蛋白的情況而修改。切割后也很容易利用其N端的HQ標(biāo)簽進(jìn)行TEV蛋白酶清除。也可以從固定在樹脂上的融合蛋白切除掉目的蛋白。
rTEV蛋白酶(重組型)
rTEV蛋白酶(重組型)是經(jīng)過基因工程改造后的重組蛋白酶,該酶特異性識Glu-Asn-Leu-Tyr-Phe- Gln-Gly七氨基酸序列。rTEV蛋白酶與腸激酶U(EK)、Thrombin、FactorXa、SUMO等蛋白酶相比,具有高活性、高特異性的雙重特點(diǎn),rTEV蛋白酶因具高剪切活性和特異性,已成為融合蛋白表達(dá)后去除融合tag的蛋白酶。該酶經(jīng)6×His標(biāo)簽純化而得(含組胺酸標(biāo)簽),純度達(dá)99%,剪切反應(yīng)完畢后可通過His標(biāo)簽純化樹脂Ni-NTA Resin(Cat.No. P2010)去除。該酶在2-8℃-30℃溫度、pH范圍(6.0-8.5)反應(yīng)條件下均具有活性(見下表)
不同溫度下剪切活性(%)
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
組分與保存條件:
貨號 組分名稱 規(guī)格 數(shù)量 保存
P2060 rTEV 蛋白酶 (凍干粉) 1000U 1支 -70℃/-20℃
P2060-A rTEV Buffer A 1 ml 1支 -20℃
P2060-B 500 X rTEV Buffer B 1ml 1支 -20℃
備注:rTEV 蛋白酶保存條件:長期儲存-70℃,可儲存1年,-20℃,可儲存6個月。
剪切活性(%)電泳圖譜
使用方法說明:
1. 推薦使用溶液:50 mM NaH2PO4, 500mM NaCl, 1mM EDTA, 1mMDTT, 10%甘油,pH 8.0 buffer中進(jìn)行。
2. rTEV 蛋白酶 (凍干粉)1000U/支用1ml rTEV Buffer A溶解。
3. rTEV與需要剪切的蛋白比例:1:100
4. 推薦剪切時間僅供參考:用戶可以根據(jù)自己研究的蛋白進(jìn)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
別名:TEV protease
表達(dá)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
種屬:Human
表達(dá)宿主: E.Coli
表達(dá)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
種屬:Human
表達(dá)宿主: E.Coli
純度:> 95%, as determined by SDS-PAGE
內(nèi)毒素:< ﹤1.0 EU per 1μg cytokine as determined by the LAL method.
穩(wěn)定性:Samples are stable for up to twelve months from date of receipt -70°C
預(yù)測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.
緩沖液: Supplied as a 0.2μm filtered solution of 50mM Tris, 10%Sucrose,5mM DTT pH8.0
SDS-PAGE:
內(nèi)毒素:< ﹤1.0 EU per 1μg cytokine as determined by the LAL method.
穩(wěn)定性:Samples are stable for up to twelve months from date of receipt -70°C
預(yù)測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.
緩沖液: Supplied as a 0.2μm filtered solution of 50mM Tris, 10%Sucrose,5mM DTT pH8.0
SDS-PAGE:
Usage Guide
儲存方法: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.
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.