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TobaccoEtchVirus(TEV)proteaseisacysteineproteasethatiscommonlyusedtoremovefusiontags,includingHis,GSTandMBP,attheNorCterminiofrecombinantproteins.TEVproteasespecificallyrecognizesasevenaminoacidsequenceGlu-Asn-Leu-Tyr-Phe-Gln-Gly/Ser(ENLYFQG/S)andcleavesbetweenGlnandGly/Serunderawiderangeofconditions.
Recombinant6xHis-TEVwasexpressedinE.Coliandpurifiedtonearhomogeneity.ASer219Asn(S219N)substitutionwasintroducedthathasnoeffectonTEVproteaseactivity,butinhibitsauto-cleavageandthus,stABIlizingTEVproteaseduringincubationandlong-termstorage.
Werecommendusing3%suppliedTEVprotease(w/w)topreformreactionsat40Cfor12-16horat300Cfor1-4h.Dependingonspecificsubstrates,TEVproteaseconcentration,incubationtime,andreactiontemperatureshouldbeoptimized.6xHis-TEVcanberemovedbyNi-NTAresinafterincubation.
TEVproteasemaintainshighactivityunderawiderangeoftemperatures(40C-300C),pH(6.0-8.5)andbuffers,andinthepresenceofcommonlyusedproteaseinhibitors.
Recombinant6xHis-TEVwasexpressedinE.Coliandpurifiedtonearhomogeneity.ASer219Asn(S219N)substitutionwasintroducedthathasnoeffectonTEVproteaseactivity,butinhibitsauto-cleavageandthus,stABIlizingTEVproteaseduringincubationandlong-termstorage.
Werecommendusing3%suppliedTEVprotease(w/w)topreformreactionsat40Cfor12-16horat300Cfor1-4h.Dependingonspecificsubstrates,TEVproteaseconcentration,incubationtime,andreactiontemperatureshouldbeoptimized.6xHis-TEVcanberemovedbyNi-NTAresinafterincubation.
TEVproteasemaintainshighactivityunderawiderangeoftemperatures(40C-300C),pH(6.0-8.5)andbuffers,andinthepresenceofcommonlyusedproteaseinhibitors.
AdditionalInformation
ProductName: | 6xHis-TEVProtease |
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AlsoKnownAs: | TEVprotease,NIa,Tobaccoetchvirusprotease |
CatalogNo.: | P1001-200 |
Size: | 200µg |
MolecularWeight: | 27kDa |
Species: | TabaccoEtchVirus |
Source: | Bacterialrecombinant |
Stock: | 100mMTris,pH8.5at40C,500mMNaCl,5mMDTT,0.5mMEDTAand50%Glycerol |
Concentration: | Seetubelabel |
QualityAssurance: | >90%puritybasedonSDS-PAGE. |
Storage: | Storeat-80°C;avoidmultiplefreeze-thawcycles |
PDFDataSheet: | PDFDatasheet,MSDS |
NCBIRefSeq: | N/A |
Image(s): | ![]() |
ShippingMethod: | Dryiceshipping |
References: | 1.LucastLJ.,etal.(2001)Biotechniques30,544. |