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SelecTEV™Proteaseisa27kDimprovedformofTobaccoEtchVirus(TEV)proteasethathasbeenengineeredtobemorestable,active,andspecificthanthenativeprotease.SelecTEV™ProteaseisacysteineproteasethatrecognizestheTEVrecognitionsequenceGlu-Asn-Leu-Tyr-Phe-Gln-GlyandcleavesbetweenGlnandGly.Thisseven-amino-acidsequenceisrarelyfoundinproteins,makingSelecTEV™Proteaseanidealchoicefortagremovaloffusionproteins.Theoptimaltemperatureforcleavageis30°C,howevertheenzymeisactiveoverawidetemperaturerange(4–30°C)andpH(6.0–8.5)toaccommodateyourspecificprotein.Followingdigestion,SelecTEV™ProteaseiseasilyremovedfromthecleavagereactionbyaffinitychromatographyusingthepolyhistidinetagattheN-terminusoftheprotease.SelecTEV™ProteaseispurifiedfromE.colibyaffinitychromatographyusingthepolyhistidinetagandis90%purewhenvisualizedonanSDS-PAGEgel.
SelecTEVProteasecanbeusedwiththeExpressoSolubilityandExpressionScreeningSystemvectors,whichcontainaTEVcleavagerecognitionsite.
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Coomassie-stainedSDS-PAGEgelofTEVcleavagereaction.10UofSelecTEVProteasewasincubatedwith30µgoffusionproteinsubstrateat30°Cfor1hourina100µLreaction.Marker(laneM),SelecTEVProtease(Lane1),Uncleavedfusionsubstrate(Lane2),CleavedsubstrateandSelecTEVProtease.(Lane3) |
30810-1:SelecTEVProtease(1tube),100mMDTT(1tube),20XSelecTEVBuffer(1tube)
30810-2:SelecTEVProtease(5tubes),100mMDTT(5tubes),20XSelecTEVBuffer(5tubes)