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MultiplexyourproteinsolubilityscreenOneconvenientscreeningkitcontaining7fusiontagsproventoincreaseproteinexpressionandsolubilityUseoneamplicontosimultaneouslygenerateallyourconstructsRapid,simultaneousproteinscreeningwithTEV-cleavablefusiontagsPCRtoproteininaslittleas4daysOverviewSimplifiedworkflowResultsFusiontagsincludedProteincleavagewithSelecTEV™ProteaseFAQ'sOverviewTheExpresso®SolubilityandExpressionScreeningSystemisanall-in-onesystemthatenablesPCRamplification,rapidcloning,andexpressionofyourproteinofinterestwithadiversepanelofpowerfulandcleavablesolubilitytags.Lucigenhasidentified7uniqueandliterature-validatedsolubilitytagsandcombinedthemwithExpressioneering™technologytocreateasuiteofcloning-readyvectorsforrapidandparallelevaluationofyourproteinwithall7solubilitytags(plusa6xHis-taggedcontrolvector).Figure1.EachpSOLvectorcontainseverythingyouneedtoseamlesslycloneinyourtargetgenewithoutanyadditionalaminoacidsadded. FusiontagsarecleavableviaaTEVproteasesite. Backtotop▲SimplifiedWorkflowLucigen'sExpressioneering™technologymakesitsimplerthanevertorapidlyscreenmultiplefusiontagsforsolubilityandexpressionenhancement. Useoneamplicontosimultaneouslycreate8differentclones(7fusiontagsandoneHis-taggedcontrol),followedbyscreeningforsolubilityandexpression.WanttolearnmoreaboutExpressioneering? WatchourvideoBacktotop▲ResultsEveryproteinreactsdifferentlytovariousfusiontags. WiththeExpressoSolubilityandExpressionScreeningSystem,youcanquicklyandeasilyscreenmultiple,provenfusiontagstofindtheoptimaltagforyourprotein.Figure2.GH1(HumanGrowthHormone)wasevaluatedforE.coliexpressionandsolubilitywhenexpressedasafusion.FusiontagsSlyD,Tsf,SUMO,Bla,andMBPdemonstratedenhancedsolubility(LaneS)comparedtotheHis-controlprotein.T:TotalLysate,S:ClearedSolubleFraction,I:InsolublePelletBacktotop▲FusionTagsIncludedTheExpressoSolubilityandExpressionScreeningSystemincludes7vectorsthateachcontainauniquefusiontagandoneHiscontrolvector.Thefusiontagsvaryinsequence,size,andderivative.Belowisatablesummarizingthecharacteristicsofeachfusiontag:#FusionTagAALengthkDa*pIDescription16xHis-AFV11313.55.0HypotheticalproteinfromAcidianusfilamentousvirus126xHis-SlyD21022.75.2FKBP-typepeptidyl-prolylcis-transisomerase36xHis-Tsf29732.25.7E.colielongationfactor46xHis-SUMO11513.35.2SmallUbiquitin-likeModifier56xHis-Bla38141.34.4Beta-lactamase66xHis-MBP38242.15.5Maltose-BindingProtein76xHis-GST23327.46.6GlutathioneS-transferase86xHisControl141.87.0AffinitytagBacktotop▲CleavagewithSelecTEV®ProteaseAllfusiontaggedproteinsproducedwiththeExpressoSolubilityandExpressionScreeningSystemarefullycleavablewithSelecTEVproteasefromLucigen. Formoreinformation,visittheSelecTEVproteaseproductpageFigure3.Coomassie-stainedSDS-PAGEgelofTEVcleavagereaction.10UofSelecTEVProteasewasincubatedwith30µgoffusionproteinsubstrateat30°Cfor1hourina100µLreaction.Marker(laneM),SelecTEVProtease(Lane1),Uncleavedfusionsubstrate(Lane2),CleavedsubstrateandSelecTEVProtease.(Lane3)Backtotop▲ORDERINFORMATIONTheExpressoSolubilityandExpressionScreeningSystemcontainspre-processedpSol™vectorswith7differentfusiontagsandoneHIS-taggedcontrolvector(3reactionsofeachvector),positivecontrolinsert,sequencingprimers,E.cloni®10GChemicallyCompetentCells(SOLOs),and20%Rhamnosesolutionand15%Glucosesolutionautoinductionreagents.SelecTEV™ProteaseandAccura®High-FidelityPolymeraseareavailableforpurchaseseparately.Forresearchuseonly.Completeprotocolsareavailableonlineatwww.lucigen.com/manuals.
美国Lucigen公司,自1998年成立至今,一直致力于生命科学领域相关科研产品的研究与开发,在分子生物学领域处于领导性地位。Lucigen公司主要开发各类用于基因克隆的试剂盒及相关产品,包括:CloneSmart®平端克隆试剂盒、BigEasy®线性克隆系统、pEZSeq™平端克隆试剂盒、ClonePlex™ AK文库构建试剂盒、DNA Terminator ®末端修复试剂盒、EconoTaq® DNA聚合酶及E.cloni感受态细胞等。Lucigen公司凭借其独到的产品技术,过硬的产品质量,良好的产品服务赢得了全球广大用户的信赖。
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Biosearch technologies lucigen感受态细胞选择指南_北京中北林格...
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Lucigen/EconoTaq DNA Polymerase (separate Mg++)/30032-2/5,000 U (5 x 1,000 U)
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Lucigen/DNATerminator® End Repair Kit/40035-2/50 rxns
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Lucigen/E. cloni 10G ELITE Electrocompetent Cells > 2 × 10^10 cfu/µg/60052-1/12 rxns (DUOs)
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Lucigen/FailSafe™ PCR 2X PreMix L/FSP995L/2.5 ml