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

================  蚂蚁淘在线  ================

免责声明：本文仅代表作者个人观点，与本网无关。其创作性以及文中陈述文字和内容未经本站证实，对本文以及其中全部或者部分内容、文字的真实性、完整性、及时性本站不做任何保证或承诺，请读者仅作参考，并请自行核实相关内容

版权声明：未经蚂蚁淘在线授权不得转载、摘编或利用其他方式使用上述作品。已经经本网授权使用作品的，应该授权范围内使用，并注明“来源：蚂蚁淘在线”。违反上述声明者，本网将追究其相关法律责任。