$100DiscountCodeforYourFirstOrderof1mgCreRecombinase:CreDiscount

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Crerecombinase,oftenabbreviatedtoCre,isaTypeItopoisomerasefromP1bacteriophagethatcatalyzessite-specificrecombinationofDNAbetweenloxPsites.Thisenzymedoesnotrequireanyenergycofactors,andCre-mediatedrecombinationquicklyreachesequilibriumbetweensubstrateandreactionproducts.TheloxPrecognitionelementisa34basepair(bp)sequencecomposedoftwo13bpinvertedrepeatsflankingan8bpspacerregionwhichconfersdirectionality.RecombinationproductsaredependentonthelocationandrelativeorientationoftheloxPsites.TwoDNAspeciescontainingsingleloxPsiteswillbefusedwhilstDNAbetweenloxPsitesinthesameorientationwillbeexcisedincircularformandDNAbetweenopposingloxPsiteswillbeinvertedwithrespecttotherestoftheDNA.

Crerecombinaseisusedasatooltomodifygenesandchromosomes.InthisapproachtheCrerecombinaseisusedtodeleteasegmentofDNAflankedbyLoxPsites(aka'floxed')inanexperimentalanimal.Ithasbeenusedtogenerateanimalswithmutationslimitedtocertaincelltypes(tissue-specificknockout)oranimalswithmutationsthatcanbeactivatedbydrugadmiNISTration(inducIBLeknockout)inanumberoftransgenicspecies.TheavailABIlityoftransgeniclineswithtissuespecificorinducibleCreexpressionpermitsresearcherstoinactivateoractivateageneofinterestsimplybybreedingafloxedanimaltopre-existingCre-transgenics.OneexampleofaninducibleCrerecombinasesystemistheCre-ER(ER=EstrogenReceptor)systeminwhichintraperitonealinjectionoftamoxifenwillcausedose-dependentexcisionofthefloxedsite(i.e.willinactivatethegeneofchoice).

RecombinantCrerecombinase(TAT-Cre)waspurifiedfromanE.colistraincarryinganengineeredplasmidencodingenhancedformofCreRecombinasefrombacteriophageP1.ThisCrerecombinasehasanN-terminal6XHistag,aTatpeptide(GRKKRRQRRRPPAGTSVSL)andanNLSsequence(PKKKRKV).HTNCisthemosteffectiveproteinintransduction(invivo)andsubsequentrecombinationcomparedtootherformsofCrerecombinases,e.g.,HNC,TCH6,HC,HNCM,CH.Incubationoffibroblastreportercellswith1μMHTNCfor1to2hourscanresultintranductionof60~90%ofthecells*.Additionof100μMchoroquinetoculturemediumcanfurtherenhancetransductionandrecombination.

*Usethispagetocalculateconcentrationrequiredforculturedcells: http://www.excellgen.com/pub/mw_2_moles.html

• InvitroLoxPrecombinationforsubcloningorvector/cloneengineering

• TransductionintoculturedcellsincludingstemcellsexVivo

MolecularWeight:43kDa

QC:Tat-Creinduced80~100%recombinationefficienciesinHEK293T-Crereportercells.HEK293T-cellsweretransfectedwithreporterplasmidDNA:LoxP-RFP-Stop-LoxP-GFP,thenadd1μMpurifiedTat-Creprotein.

Fig1:0:beforeaddingTat-Cre;1to5:1to5daysaftertransductionwithTat-Cre.

Fig2:ComparisonofrecombinationacitivitiesofTat-Crerecombinaseinliquidand lyophilizedform.1,Tat-Crerecombinaseinliquidformafterstoringat-80oCfor6months;2,LyophilizedTat-Crerecombinaseafterstoringat37oCfor2weeks;3,LyophilizedTat-Crerecombinaseafterstoringat37^oCfor2weeksandsubsequentlystoringat4^oCfor2weeks.Picturesweretaking15hoursafteradding1uMTat-Crerecombinase.

EndotoxinLevels:<0.1EU/μg

Purity:>98%bySDS-PAGEandHPLCanalysis.

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