Description | FLPrecombinasecatalyzesthesite-specificrecombinationofDNAbetweenFRTsites.Ithasbeenwidelyusedforgeneratingconditionalknockoutmice,mediatedbyFLP/FRTsystem. Recombinantcell-permeantFLPrecombinaseTat-FlpwaspurifiedfromanHEK293celllineoverexpressingFLPRecombinasefromyeastSaccharomycescerevisiaewithadditionalN-terminal6XHistag,aTatpeptide(GRKKRRQRRRPPAGTSVSL)andanNLSsequence(PKKKRKV).Inaddition,this"Thermostable"mutantFLP(P2SL33SY108NS294P)has~4-10foldhigheractivitythanwild-typeFLPat37°C. |
Applications | exvivoFRTrecombination. |
QC | Additionof100unitsofTat-FlptoFRT-RFP-Stop-FRT-GFP293Treportercellsresultedin60~90%recombinationefficiency. Figure1:Tat-Flptransductionefficiencyisconcentrationdependent. 1:noTat-FLP;2~8:adding5,10,25,50,75,100and200unitsofTat-FLPtoHEK293TcellstransfectedwithFRT-RFP-Stop-FRT-GFPplasmidDNA.Picturesweretaken4daysaftertransductionwithvariousamountofTat-FLP |
UnitDefinition | Astandardof100UnitsisdefinedastheamountofTAT-FLPin1.0mLoftissueculturemediumthatisrequiredtoinduce50% GFPexpressioninaHEK293Treportercelllineassay. |
Source | Sf9cells |
Storage | -20~-80°C. |