Everest Biotech/TEV Protease (TurboTEV) 10 mg (100,000 Units)/1500020102/10 mg (100,000 Units),Everest Biotech,,Everest Biotech,everestbiotech,+,,Array.Array.Array蚂蚁淘厂家直采.

产品名称：Everest Biotech/TEV Protease (TurboTEV) 10 mg (100,000 Units)/1500020102/10 mg (100,000 Units)

产品编号：1500020102

市场价：¥33696.00

场地：美国(厂家直采)

产品分类：蛋白类>酶>蛋白酶类>

联系QQ：1570468124

电话号码：4000-520-616

邮箱： info@ebiomall.com

美元价：$3240.00

品牌： Everest Biotech

公司：Everest Biotech

公司分类：

商品介绍

ProteinType:
Protease

Source:
E.coli

SpecificActivity:
>10Units/µg.1UnitofTurboTEVProteasecleaves>85%of3µgofcontrolsubstratein1hourat30oC.

Formulation
2mg/mlin25mMTris-HCl,pH8.0,50mMNaCl,1mMTCEP,50%glycerol

Storage/Handling:
Storeat-20^oC

A49kDaGST-fusionprotein(C)at1mg/mlisincubatedwithTurboTEVorTEVProteaseataratioof(1)1:50,(2)1:100,(3)1:200(w/w)inabufferof25mMTris-HCl,pH8.0,150mMNaCl,14mMb-mercaptoethanolat4oCfor16hours.Thecleavedproductsare27kDaand22kDa."TEV"isacompetitors’TEVProteaseproduct.

Q)ForyourTEVdigestreaction,theprotocolstatesthatthereactionisperformedin25mMTris-HCl,pH8.0,150mMNaCland14mMβ-mercaptoethanol.IwaswonderinghowimportantthesebufferconditionsareandwhetherthepresenceofanyofthefollowingmayhaveanadverseaffectonthecleavageefficiencyofyourTurboTEV?

a)1mMDTT

b)10%Glycerol

c)20mMHistidine

d)500mMNaCl

e)100mMTris-HCl

f)Additionofβ-mercaptoethanol

a)DTTof1mMshouldbefine;

b)10%Glycerolwouldnotberecommended;

c)20mMHistidinewouldnotberecommended;

d)500mMNaClwouldnotberecommended;

e)100mMTriswouldnotberecommended;

f)β-mercaptoethanolfunctionslikeDTT,soaddingβ-mercaptoethanolshouldnotbeaproblem.

Q)Dowehavetobufferexchangemyproteinpriortocleavage?

A)Itmaybenecessary.

Step1:CleavageinSolution

MakefreshcoldDialysisBuffer

WhenmakingfreshcoldDialysisBuffer,pleasebeawarethat:

a)Thetargetproteinneedstobesolubleinthebuffer

b)ThebuffershouldNOTcontainproteaseinhibitor

c)TheDialysisBuffershouldbecompatIBLewithdownstreampurificationprocesses,e.g.minimalamountofEDTAorDTTifNicolumnwillbeusedtoremovethecleavedHis-tag.

HereisanexampleofDialysisBuffer.25mMTris-HCl,pH8.0,150-500mMNaCl,14mMb-mercaptoethanol.TurboTEVhasthesameactivityin150mMNaClor500mMNaCland400mMimidazole.

Dilutethetargetproteinpool

i.Dilutethetargetproteinpoolto1-2mg/mlwithDialysisBuffer.

Note:ThisisoptionalincasethetargetproteinaggregatesinDialysisBuffer.

ii.SaveasmallaliquotasUncutsampleforanalysis.EDTAmaybeaddedto0.5mMfinalconcentrationifthetargetproteinpooliselutedfromNicolumnandEDTAiscompatiblewiththetargetprotein.

AddTurboTEVProtease

i.AddTurboTEVProteaseataProtease:targetproteinratioof1:100(w/w)or10,000unit(1mg)TurboTEVProteaseto100mgoftargetprotein.

Note:

Thereisnoneedtocalculatethemolarratio.

TurboTEVProteasecanbeaddeddirectlytothetargetprotein.

ItisnotrequiredtochangebufferordiluteTurboTEVProtease.

Theoptimalratioshouldbedeterminedbytheuser;howeveraProtease-to-targetproteinratio(w/w)of1:50to1:200shouldworkformosttargetproteins.

Dialysis

i.DialyzeagainsttheDialysisBufferat4^oCovernight(about16hrs).ThisstepistoremoveimidazoleorglutathioneifNiorglutathionecolumnisusedtoremovethecleavedtagorTurboTEVProteaseaftercleavage.

Ifdesired,thetargetproteinpoolcanbebufferexchangedfirstbeforeTurboTEVcleavage.

Step2:RemovalofTurboTEVProtease

Applytocolumns

ThedialyzedtargetproteinandTurboTEVProteasemixturecanbeapplieddirectlytoaffinitycolumnsifcompatibleDialysisBufferisused.

ForHis-taggedprotein,useIMACtoremovethecleavedHis-tagandTurboTEVProtease.

ForGST-taggedprotein,useglutathionecolumntoremovethecleavedGST-tagandTurboTEVProtease.

Optional:SDS-PAGEanalysis

Ifdesired,analyzesamplesusingSDS-PAGEanalysis.ThedifferencebetweenthetaggedandcleavedtargetproteinmaybetoosmalltodetectbySDS-PAGE.ThecleavedHis-tagsometimescanbeseenatthebottomofthegel.

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JussiJ.Joensuu,AndrewJ.Conley,MichaelLienemann,JimE.Brandle,MarkusB.Linder,andRimaMenassa.:HydrophobinFusionsforHigh-LevelTransientProteinExpressionandPurificationinNicotianabenthamiana.PlantPhysiology,Feb2010;152:622-633.

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YoshiyukiTakatsuji,TetsuyaHaruyama,EmmaMaster,MaijaTenkanen,andMarkusB.Linder.Structure-FunctionRelationshipsinHydrophobins:ProbingtheRoleofChargedSideChains.ApplEnvironMicrobiol.Sep2013;79(18):5533-5538.

AdamBShapiro,NingGao,NicholeO¿Connell,JunHu,JasonThresher,Rong-FangGu,RossOverman,IanMHardernandGrahamGSproat.QuantitativeinvestigationoftheaffinityofhumanrespiratorysyncytialvirusphosphoproteinC-terminusbindingtonucleocapsidprotein.VirologyJournal2014,11:191

AlysM.CheatleJarvela,LisaBrubaker,AnastasiaVedenko,AnishaGupta,BruceA.Armitage,MarthaL.Nulyk,andVeroncaF.Hinman.ModularEvolutionofDNA-BindingPreferenceofaTbrainTranscriptionFactorProvidesaMechanismforModifyingGeneRegulatoryNetworks.MolBiolEvol(2014)31(10):2672-2688.

JolandaNeef,FinJ.Milder,DannyG.A.M.Koedijk,MarindyKlaassens,ErikC.Heezius,JosA.H.vanStrijp,AndreasOtto,DorteBecher,JanMaartenvanDijl,GirbeBuist.VersatilevectorsuitefortheextracytoplasmicproductionandpurificationofheterologousHis-taggedproteinsinLactococcuslactis.AppliedMicrobiologyandBiotechnology.Jul2015

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品牌介绍

Everest Biotech公司成立于2000年，由英国牛津大学一批生物化学家创建。公司科学家团队具有丰富的抗体纯化及蛋白组学专业知识，雄厚的专业背景赋予了公司能为全球科研人员提供高质量、创新性抗体产品的能力。目前，Everest Biotech可提供3000余种抗体，涉及肿瘤、自身免疫、过敏、心血管疾病、传染病、代谢、神经科学等领域。所有抗体均由Everest Biotech自主研发生产、全部经过肽亲和纯化以保证最佳的特异性和纯度、经过免疫原肽的ELISA和WB实验验证; 由PhD科学家团队提供全方位技术支持服务。Everest Biotech为众多国际知名抗体公司提供OEM服务。