Multiplexyourproteinsolubilityscreen
Oneconvenientscreeningkitcontaining7fusiontagsproventoincrease
proteinexpressionandsolubility
- Useoneamplicontosimultaneouslygenerateallyourconstructs
- Rapid,simultaneousproteinscreeningwithTEV-cleavablefusiontags
- PCRtoproteininaslittleas4days
- Overview
- Simplifiedworkflow
- Results
- Fusiontagsincluded
- ProteincleavagewithSelecTEV™Protease
- FAQ's
Overview
TheExpresso®SolubilityandExpressionScreeningSystemisanall-in-onesystemthatenablesPCRamplification,rapidcloning,andexpressionofyourproteinofinterestwithadiversepanelofpowerfulandcleavablesolubilitytags.Lucigenhasidentified7uniqueandliterature-validatedsolubilitytagsandcombinedthemwithExpressioneering™technologytocreateasuiteofcloning-readyvectorsforrapidandparallelevaluationofyourproteinwithall7solubilitytags(plusa6xHis-taggedcontrolvector).
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Figure1.EachpSOLvectorcontainseverythingyouneedtoseamlesslycloneinyourtargetgenewithoutanyadditionalaminoacidsadded. FusiontagsarecleavableviaaTEVproteasesite. |
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SimplifiedWorkflow
Lucigen'sExpressioneering™technologymakesitsimplerthanevertorapidlyscreenmultiplefusiontagsforsolubilityandexpressionenhancement. Useoneamplicontosimultaneouslycreate8differentclones(7fusiontagsandoneHis-taggedcontrol),followedbyscreeningforsolubilityandexpression.
WanttolearnmoreaboutExpressioneering? Watchourvideo
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Results
Everyproteinreactsdifferentlytovariousfusiontags. WiththeExpressoSolubilityandExpressionScreeningSystem,youcanquicklyandeasilyscreenmultiple,provenfusiontagstofindtheoptimaltagforyourprotein.
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| Figure2.GH1(HumanGrowthHormone)wasevaluatedforE.coliexpressionandsolubilitywhenexpressedasafusion.FusiontagsSlyD,Tsf,SUMO,Bla,andMBPdemonstratedenhancedsolubility(LaneS)comparedtotheHis-controlprotein.T:TotalLysate,S:ClearedSolubleFraction,I:InsolublePellet |
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FusionTagsIncluded
TheExpressoSolubilityandExpressionScreeningSystemincludes7vectorsthateachcontainauniquefusiontagandoneHiscontrolvector.Thefusiontagsvaryinsequence,size,andderivative.Belowisatablesummarizingthecharacteristicsofeachfusiontag:
# | FusionTag | AALength | kDa* | pI | Description |
1 | 6xHis-AFV | 113 | 13.5 | 5.0 | HypotheticalproteinfromAcidianusfilamentousvirus1 |
2 | 6xHis-SlyD | 210 | 22.7 | 5.2 | FKBP-typepeptidyl-prolylcis-transisomerase |
3 | 6xHis-Tsf | 297 | 32.2 | 5.7 | E.colielongationfactor |
4 | 6xHis-SUMO | 115 | 13.3 | 5.2 | SmallUbiquitin-likeModifier |
5 | 6xHis-Bla | 381 | 41.3 | 4.4 | Beta-lactamase |
6 | 6xHis-MBP | 382 | 42.1 | 5.5 | Maltose-BindingProtein |
7 | 6xHis-GST | 233 | 27.4 | 6.6 | GlutathioneS-transferase |
8 | 6xHisControl | 14 | 1.8 | 7.0 | Affinitytag |
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CleavagewithSelecTEV®Protease
AllfusiontaggedproteinsproducedwiththeExpressoSolubilityandExpressionScreeningSystemarefullycleavablewithSelecTEVproteasefromLucigen. Formoreinformation,visittheSelecTEVproteaseproductpage
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| Figure3.Coomassie-stainedSDS-PAGEgelofTEVcleavagereaction.10UofSelecTEVProteasewasincubatedwith30µgoffusionproteinsubstrateat30°Cfor1hourina100µLreaction.Marker(laneM),SelecTEVProtease(Lane1),Uncleavedfusionsubstrate(Lane2),CleavedsubstrateandSelecTEVProtease.(Lane3) |
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ORDERINFORMATION
TheExpressoSolubilityandExpressionScreeningSystemcontainspre-processedpSol™vectorswith7differentfusiontagsandoneHIS-taggedcontrolvector(3reactionsofeachvector),positivecontrolinsert,sequencingprimers,E.cloni®10GChemicallyCompetentCells(SOLOs),and20%Rhamnosesolutionand15%Glucosesolutionautoinductionreagents.SelecTEV™ProteaseandAccura®High-FidelityPolymeraseareavailableforpurchaseseparately.
Forresearchuseonly.Completeprotocolsareavailableonlineatwww.lucigen.com/manuals.
