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预优化DNA转染试剂HepG2
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发布时间：2024-12-23
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.
StorageConditionStoreat4°C.Ifstoredproperly,theproductisstablefor12monthsorlongerAPictureShowingTransfectionEfficiencyofGenJet™ReagentonHepG2Cells![\"\"](\"http://www.ebiomall.cn/images/sell/20150513171750223.jpg\")
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.![\"\"](\"http://www.ebiomall.cn/images/sell/20150513171806404.jpg\")
GenJet™reagentisoptimizedforHepG2cells(ATCC#HB-8065).HepG2cellsweregrownperATCCrecommendedculturemediumonacollagentypeItreatedculturedishandtransfectedwithGFP(pEGFP-N3,4.7kb,rightpanel)andβ-galactosidasecDNA(pSV-β-galactosidase,6.9kb,leftpanel)bypre-optimizedGenJet™DNATransfectionReagentforHepG2cells.Theefficiencywaschecked48hoursposttransfectionbyZeiss510ConfocalMicroscopyandβ-galactosidasestainingkitrespectively.![\"\"](\"http://www.ebiomall.cn/images/sell/pdf.gif\")
DataSheet.pdf
| 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% |
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本文链接： http://amaxa.immuno-online.com/view-731067.html
发布于 ： 2024-12-23
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