Forresearchuseonly.Notforuseindiagnosticprocedures.Lipofectamine®LTX&PlusReagentCat.no.Lipofectamine®LTXPlus™ReagentStorageA126210.1mL40μL4°C(donotfreeze)15338-0300.3mL0.25mL15338-1001.0mL0.85mL15338-500*15mL—*Plus™Reagent(Cat.no.11514-015)availableseparately.Pub.PartNo.15338.ppsPub.No.MAN0001225Rev.Date4October2012Description•Lipofectamine®LTXandPlus™Reagentareproprietaryformulationsfortransfectingnucleicacidsintoawiderangeofeukaryoticcells.•Nucleicacid-Lipofectamine®LTXcomplexescanbeaddeddirectlytocellsinculturemedium,inthepresenceorabsenceofserum/antibiotic.•Itisnotnecessarytoremovecomplexesorchange/addmediumaftertransfection.ImportantGuidelinesforTransfection•UseOpti-MEM®IReducedSerumMedium(Cat.no.31985-062)todiluteLipofectamine®LTXReagentandnucleicacids.•TheamountofLipofectamine®LTXReagentrequiredforsuccessfultransfectionvariesdependingonthecelltypeandpassagenumber.StartanynewtransfectionbytestingtherecommendedfourconcentrationsofLipofectamine®LTXReagenttodetermineanoptimumamount.•Foradditionalinformation,andprotocolsontransfectingsuspensioncellsrefertothemanualatwww.lifetechnologies.com/transfection5Day0Day1Day2–4CellsLipofectamine®LTXReagentDNAandPlus™ReagentSeedcellssotheyare70–90%confluentatthetimeoftransfection.Dilute4amountsofLipofectamine®LTXReagentinOpti-MEM®Medium.DiluteDNAinOpti-MEM®Medium,thenaddPlus™Reagent.AdddilutedDNAtoeachtubeofdilutedLipofectamine®LTXReagent(1:1ratio).Incubate.AddDNA-reagentcomplextocells.Visualize/analyzetransfectedcells.PlasmidTransfectionTransfectcellsaccordingtothefollowingtable.UsetheindicatedvolumeofDNAandPlus™ReagentwitheachofthefourvolumesofLipofectamine®LTX.Eachreactionmixissufficientfortriplicate(96-well),duplicate(24-well),andsinglewell(6-well)transfections,andaccountforpipettingvariations.Forevenlesstoxicity,reducetheamountofDNA-reagentcomplextothecells,orreducetheamountofDNAusedtomakecomplexes.Component96-well24-well6-wellAdherentcells1–4x1040.5–2x1050.25–1x106Opti-MEM®Medium25...
