Everest Biotech/HRV 3C Protease 1 mg (1,000 Units)/1500010012/1 mg (1,000 Units),Everest Biotech,,Everest Biotech,everestbiotech,+,,Array.Array.Array蚂蚁淘厂家直采.

产品名称：Everest Biotech/HRV 3C Protease 1 mg (1,000 Units)/1500010012/1 mg (1,000 Units)

产品编号：1500010012

市场价：¥2714.40

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：$261.00

品牌： Everest Biotech

公司：Everest Biotech

公司分类：

商品介绍

ProteinType:
Protease

Source:
E.coli

SpecificActivity:
>1Units/µg.1UnitsofTurbo3C(HRV3C)Proteasecleaves>95%of100µgoftargetproteinat4^oCfor16hours.

Storage/Handling:
Storeat-80^oC.

2mg/mlin50mMTris-HCl,pH8.0,150mMNaCl,1mMEDTA,1mMTCEP,50%glycerol

A68kDaGST-fusionprotein(C)at1mg/mlisincubatedwithTurbo3C(HRV3C)Protease(*)ataratioof(1)1:50,(2)1:100,(3)1:200,(4)1:400(w/w)inabufferof25mMTris-HCl,pH8.0,150mMNaCl,14mMb-mercaptoethanolat4^oCfor16hours.Thecleavedproductsare42kDaand26kDa

CleavageinSolution

MakefreshcoldDialysisBuffer.DialysisBuffershouldbeabufferinwhichthetargetproteinissoluble.ThereshouldbenoproteaseinhibitorintheDialysisBuffer.TheDialysisBuffershouldbecompatIBLewithdownstreampurificationprocesses,e.g.minimalamountofEDTAorDTTifNicolumnwillbeusedtoremovethecleavedHis-tag.
HereisanexampleofDialysisBuffer.25mMTris-HCl,pH8.0,150-500mMNaCl,14mMb-mercaptoethanol
Turbo3Chasthesameactivityin150mMNaClor500mMNaCland400mMimidazole.
Dilutetheproteinpoolto1-2mg/mlwithDialysisBuffer.ThisisoptionalincasethetargetproteinaggregatesinDialysisBuffer.SaveasmallaliquotasUncutsampleforanalysis.EDTAmaybeaddedto0.5mMfinalconcentrationifthetargetproteinpooliselutedfromNicolumnandEDTAiscompatiblewiththetargetprotein.
AddTurbo3CProteaseataProtease:targetproteinratioof1:100(w/w)or1,000unitTurbo3CProteaseto100mgoftargetprotein.Thereisnoneedtocalculatethemolarratio.Turbo3CProteasecanbeaddeddirectlytothetargetprotein.ThereisnoneedtochangebufferordiluteTurbo3CProtease.Theoptimalratioshouldbedeterminedempirically.AProtease-to-targetproteinratio(w/w)of1:50to1:200shouldworkformosttargetproteins.
DialyzeagainsttheDialysisBufferat4^oCovernight(about16hrs).DialysisistoremoveimidazoleorglutathioneifNiorglutathionecolumnisusedtoremovethecleavedtagorTurbo3CProteaseaftercleavage.Ifdesired,thetargetproteinpoolcanbebufferexchangedfirstbeforeRurbo3Ccleavage.
RemovalofTurbo3CProtease
1. ThedialyzedtargetproteinandTurbo3CProteasemixturecanbeapplieddirectlytoaffinitycolumnsifcompatibleDialysisBufferisused.ForHis-taggedprotein,useIMACtoremovethecleavedHis-tagandTurbo3CProtease.ForGST-taggedprotein,useglutathionecolumntoremovethecleavedGST-tagandTurbo3CProtease.
2. Ifdesired,analyzesamplesusingSDS-PAGEanalysis.ThedifferencebetweenthetaggedandcleavedtargetproteinmaybetoosmalltodetectbySDS-PAGE.ThecleavedHis-tagsometimescanbeseenatthebottomofthegel.