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预优化DNA转染试剂HepG2
来自 : 发布时间:2024-05-03

DescriptionGenJet™DNAInVitroTranfectionReagentforHepG2Cellsispre-optimizedforHepG2celltransfection.HepG2(Hepatocellularcarcinoma,human)isaperpetualaddcelllinewhichwasderivedfromthelivertissueofa15yearoldcaucasianmalewithawelldifferentiatedhepatocellularcarcinoma.Thesecellsareepithelialinmorphology,haveamodelchromosomenumberof55andarenottumorigenicinnudemice.Thecellssecreteavarietyofmajorplasmaproteinse.g..albumin,alpha2-macroglobulin,alpha1-antitrypsin,transferrinandplasminogen.Theyhavebeengrownsuccessfullyinlargescalecultivationsystems.HepatitisBvirussurfaceantigenshavenotbeendetected.HepG2cellshavebeenshowntobeG418resistant(400µg/mL).Thecellswillrespondtostimulationwithhumangrowthhormone.RefertothefollowingoptimaltransfectionconditionsformaximaltransfectionefficiencyonHepG2cells.GenJet™reagent,1.0ml,issufficientfor300to600transfectionsin24wellplatesor150to300transfectionsin6wellplates.
SummaryofOptimalTransfectionConditions:HepG2cellcultureConfluenceonthedayoftransfectionCellcultureconditionsGenJet™(µl):DNA(µg)RatioDiluentforDNAandTransfectionReagentIncubationTimetoFormGenJet™/DNAComplexPresenceofSerum/AntibioticsduringTransfectionChangeMedium5HoursAfterTransfectionMaximalEfficiencyTransfectionResults:ReporterGenePlasmidEfficiency(GFP%)collagentypeIpre-treatedculturedish~90%DMEMwith4.5g/Lglucose,10%FBS3:1Serum-freeDMEMwith4.5g/Lglucose15minutesatRTYesNo48hoursEGFPpEGFP-N3(CMVpromoter)82%
StorageConditionStoreat4°C.Ifstoredproperly,theproductisstablefor12monthsorlongerAPictureShowingTransfectionEfficiencyofGenJet™ReagentonHepG2Cells\"\"GenJet™reagentisoptimizedforHepG2cells(ATCC#HB-8065)withexceptionalefficiencyincomparisonofLipofectamine2000(L2K)andAmaxaelectroporationdevice.HepG2cellsweregrownperATCCrecommendedculturemediumonacollagentypeItreatedculturedishandtransfectedwithpEGFP-N3byGenJet™.Theefficiencywaschecked48hoursposttransfection.RightPanel:ComparisonoftransfectionefficiencyofGenJetwithLipofecatmine2000(L2K),andAmaxaelectroporationdeviceonHepG2cells.GFPDNA(pEGFP-N3,4.7kb)wasintroducedtoHepG2cell(culturedonCollagenpretreateddishes)withdifferenttransfectionreagentspermanufacturersprotocols.GFPpositivecell(%)andfluorescenceintensityweredetectedbypassingthroughFACS48hoursposttransfectionLeftPanel:presenceofserumandantibioticsenhancesGenJetreagentsefficiencyonHepG2cells.HepG2cell(grownoncollagentreateddishes)wastransfectedwiththreedifferentconditions-------serumandantibioticsfree,presenceof10%serumandantibioticsfollowedbyremoval5hoursposttransfectionandpresenceof10%serumandantibioticswithoutremoval5hoursposttransfection.\"\"GenJet™reagentisoptimizedforHepG2cells(ATCC#HB-8065).HepG2cellsweregrownperATCCrecommendedculturemediumonacollagentypeItreatedculturedishandtransfectedwithGFP(pEGFP-N3,4.7kb,rightpanel)andβ-galactosidasecDNA(pSV-β-galactosidase,6.9kb,leftpanel)bypre-optimizedGenJet™DNATransfectionReagentforHepG2cells.Theefficiencywaschecked48hoursposttransfectionbyZeiss510ConfocalMicroscopyandβ-galactosidasestainingkitrespectively.\"\"DataSheet.pdf

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发布于 : 2024-05-03 阅读()