预优化DNA转染试剂HepG2相关交流-蚂蚁淘在线

预优化DNA转染试剂HepG2

mayitao /发布时间:1629135002 /浏览量:77

Description

GenJet™DNA InVitro TranfectionReagentforHepG2Cellsispre-optimizedforHepG2 celltransfection. 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/AntibioticsduringTransfectionChangeMedium5HoursAfterTransfectionMaximalEfficiency TransfectionResults: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), and Amaxaelectroporationdeviceon HepG2cells.GFPDNA(pEGFP-N3,4.7kb)wasintroducedtoHepG2cell(culturedonCollagenpretreateddishes)withdifferenttransfectionreagentspermanufacturersprotocols.GFPpositivecell(%)andfluorescenceintensityweredetectedbypassingthroughFACS48hoursposttransfection LeftPanel: presenceofserumandantibioticsenhancesGenJetreagentsefficiencyonHepG2cells.HepG2cell(grownoncollagentreateddishes)wastransfectedwiththreedifferentconditions-------serumandantibioticsfree,presenceof10%serumandantibioticsfollowedbyremoval5hoursposttransfectionandpresenceof10%serumandantibioticswithoutremoval5hoursposttransfection.

GenJet™reagentisoptimizedforHepG2cells(ATCC#HB-8065). HepG2cellsweregrownperATCCrecommendedculturemediumonacollagentypeItreatedculturedishandtransfectedwithGFP(pEGFP-N3,4.7kb,rightpanel)and β-galactosidasecDNA(pSV-β-galactosidase,6.9kb,leftpanel)bypre-optimizedGenJet™DNATransfectionReagentforHepG2cells.Theefficiencywaschecked48hoursposttransfectionbyZeiss510ConfocalMicroscopyand β-galactosidasestainingkitrespectively.

DataSheet.pdf

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