$100DiscountCodeforYourFirstOrderof1mgCreRecombinase:CreDiscount

Discountforswitchingfromcompetitors'product:$150.EmailtoTechSupportts@Excellgen.comfordiscountcode.

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*.Additionof100μMchoroquinetoculturemediummay furtherenhancetransductionandrecombination.

*Usethispagetocalculateconcentrationrequiredforcultured cells: 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.

EndotoxinLevels:<0.1EU/μg

Purity:>98%bySDS-PAGEandHPLCanalysis.

• 1.Integrin-LinkedKinaseRegulatesProcessExtensioninOligodendrocytesviaControlofActinCytoskeletalDynamics,TheJournalofNeuroscience,5June2013,33(23):9781-9793;doi:10.1523/​JNEUROSCI.5582-12.2013
• 2.CuttingEdge:Krüppel-likeFactor2IsRequiredforPhenotypicMaintenancebutNotDevelopmentofB1BCells.TheJournalofImmunology. August31,20121201439.doi:10.4049/​jimmunol.1201439.http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3448866/
• 3.KINASE-INDEPENDENTFEEDBACKOFTHETAK1/TAB1COMPLEXONBCL10TURNOVERANDNF-κBACTIVATION.Mol.Cell.Biol.2013.doi:10.1128/MCB.06407-11
• 4.Defectiveglucosemetabolisminpolycystickidneydiseaseidentifiesanewtherapeuticstrategy.NatureMedicine19,488–493(2013)doi:10.1038/nm.3092
• 5.TCell−DerivedIL-17MediatesEpithelialChangesintheAirwayandDrivesPulmonaryNeutrophilia.JImmunolpublishedonline21August2013http://www.jimmunol.org/content/early/2013/08/21/jimmunol.1301360
• 6.RapidconversionofEUCOMM/KOMP-CSDallelesinmouseembryosusingacell-permeableCrerecombinase.TransgenicResearch.February2014,Volume23,Issue1,pp177-185.doi:10.1007/s11248-013-9764-xNCBI:http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3890051/
• 7.Antibody-membraneswitch(AMS)technologyforfacilecelllinedevelopment. ProteinEngineering,DesignandSelection(2014)27(10):309-315.doi:10.1093/protein/gzu039
• 8.TheroleofantisenselongnoncodingRNAinsmallRNA-triggeredgeneactivation.RNA.October24,2014.doi:10.1261/rna.043968.113
• 9.ASingleOncogenicEnhancerRearrangementCausesConcomitantEVI1andGATA2DeregulationinLeukemia.Cell.Volume157,Issue2,p369–381,10April2014.DOI:10.1016/j.cell.2014.02.019