Description
Byutilizingourinnovativeandproprietarylipidconjugationtechnology,LipoD293(Ver.II)isspeciallydesignedandformulatedbyaddingproprietaryenhancersfortransfectingHEK293cellsandothermammaliancells(Figure1).Asa2ndgenerationliposomebasedDNAtransfectionreagent,LipoD293(Ver.II)offersextremelyhightransfectionefficienciesforHEK293relatedcellsaswellasmanymammaliancellswithlesscytotoxicity.LipoD293reagent(Ver.II),upgradedfromitspreviousversionwithrefinedchemistry,is3~4timesmoreefficientingenedelivery.LipoD293reagent(Ver.II),1.0ml,issufficientfor~666transfectionsin24wellplatesor~333transfectionsin6wellplates.
Figure1.ACartoonShowingLipoD293(Ver.II)EnhancedGeneDelivery
Features
-Topchoiceforhard-to-transfectcells
-Exceptionalhightitersofvirusproduction
-EquallygoodforverylongDNAs(upto180kb)
-EquallygoodforsUSPension293cells(e.g.,293F,293H,etc)
-Highlevelsofrecombinantproteinproduction
-Presenceofserumandantibioticsenhancesefficiencyon293cells
-ExceptionalefficiencyforbothsingleDNAtransfectionandmultiDNAsco-transfection
-Veryaffordable
StorageCondition
Storeat4°C.Ifstoredproperly,theproductisstablefor12monthsorlonger.
ComparisonsofTransfectionEfficiencyofLipoD293DNAInVitroTransfectionReagent(Ver.II)withBrandNameProducts
ComparisonoftransfectionefficiencyofLipoD293reagent(Ver.II)vs.lipofectamine2000(L2K)andFugeneHDonHepG2cells.
RightPanel:ComparisonoftransfectionefficiencyofLipoD293(Ver.II)withLipofecatmine2000(L2K),andFugeneHDonHepG2cells.GFPDNA(pEGFP-N3)wastransfectedwithdifferenttransfectionreagentspermanufacturer"sprotocolstoHepG2cell(culturedonCollagenpretreateddishes).GFPpositivecell(%)andfluorescenceintensityweredetectedbypassingthroughFACS48hoursposttransfection
LeftPanel:presenceofserumandantibioticsenhancesLipoD293(Ver.II)efficiencyonHepG2cells.HepG2cell(grownoncollagentreateddishes)wastransfectedwiththreedifferentconditions-------serumandantibioticsfree,presenceof10%serumandantibioticsfollowedbyremoval5hoursposttransfectionandpresenceof10%serumandantibioticswithoutremoval5hoursposttransfection.
ComparisonoftransfectionefficiencyofLipoD293reagent(Ver.II)vs.lipofectamine2000(L2K),TransITandFugene6onCHOcells.
RightPanel:ComparisonoftransfectionefficiencyofLipoD293(Ver.II)withLipofecatmine2000(L2K),TransITandFugene6onCHOcells.DNAsencodingRenillaluciferase(phRL-CMV)andGFP(pEGFP-N3)weretransfectedwithdifferentDNAtransfectionreagentpermanufacturer"sprotocols.RenillaluciferaseactivityandGFPfluorescenceweredetectedwithRenillaAssaySystemandaNikonEclipsefluorescentmicrocopyrespectively24hoursposttransfection.
LeftPanel:Comparisonofprice($/1.0mlvial)ofLipoD293versusthoseofLipofecatmine2000(L2K),TransITandFugene6.Allthepriceswerecollectedfromthemanufacturers"websites.
AcomparisonoftransfectionefficiencyofLipoD293reagentwithlipofectamine2000(L2K)onahard-to-transfectcell,primaryrataorticsmoothmusclecells.TherataorticsmoothmusclecellswerepreparedandtransfectedwithpEGFP-N3byLipoD293reagent(leftpanel)andLipofecatmine2000(L2K,rightpanel)respectivelypermanufacturers"protocols.ThetransfectionefficiencywasevaluatedbydetectingGFPfluorescencewithaNikonEclipse2000microscopy24hoursposttransfection.TheabovepictureswerekindlyprovidedbyDr.NickolaiDulinofSectionofPulmonaryandCriticalCare,UniversityofChicago.
AcomparisonoftransfectionefficiencyofLipoD293reagentwithFugeneHDonhard-to-transfectcell,LNCapcells.TheLNCapcellsweregrownasATCCrecommendedproceduresandco-transfectedwithpBabe-hygro-SSeCKs(1.5ug)andpEGFP-N3(0.5ug)perwell(6wellplate)LipoD293reagent(leftpanel)andFugeneHD(rightpanel)respectivelypermanufacturers"protocols.ThetransfectionefficiencywasevaluatedbydetectingGFPfluorescencewithaNikonEclipse2000microscopy24hoursposttransfection.TheabovepictureswerekindlyprovidedbyDr.LynGaoofRoswellParkCancerInstitute.
TwoexamplesshowingexceptionalefficiencyofLipoD293reagentonhard-to-transfectcellslikeHepG2andSaoS-2cells.HepG2andSaoS-2cellsin95%confluencyweretransfectedwithpEGFP-N3andpSV-?-galactosidaseDNAsrespectivelyinpresenceofserum/antibiotics.Theefficiencywaschecked48hoursposttransfectionbyZeiss510ConfocalMicroscopyand?-galactosidasestainingkitrespectively.
AcomparisonoftransfectionefficiencyofLipoD293reagentwith293fectinonHEK293cells.HEK293cellstransfectedwithpEGFP-C1plasmidusingLipoD293InVitroDNATransfectionReagent(Ver.II)(upperpanel)andthemostpopularbrandproduct293fectinofInvitrogen(lowerpanel).ThecellswerevisualizedbyNikonEclipseFluorescencemicroscopewithDICphaseimaging(leftpanel)andFITCimaging(rightpanel)24hourspost-transfection.
AcomparisonofLipoD293reagentvs.293fectin,XfectandFugene6transfectionreagentsonproteinproductionwithsuspension293Fcells.30mLof293FcellculturedinstandardculturemediumwastransfectedwithpEGFP-6xHisplasmidusingLipoD293InVitroDNATransfectionReagent(Ver.II)(20ugplasmidDNA),293Fectin(30ugplasmidDNA),Xfect(30ugplasmidDNA)andFugene6(30ugplasmidDNA)permanufacturers"standardtransfectionprotocols. GFPfluorescencewasvisualized48hoursposttransfection(leftpanel)withAforLipoD293,Bfor293fectin,CforXfectandDforFugene6. The6xHistaggedGFPproteinwasthenpurifiedviaNi-NTAaffinitycolumn.5ulof1stelutionfractionwasresolvedonSDS-PAGEfollowedbyCoomassieBrilliantBluestaining(rightupperpanelE)withthelane1forLipoD293,lane2forproteinMarker,lane3forXfect,lane4for293fectinandlane5forFugene6. Theproteinyieldwasquantifiedviaspectrometer(rightlowerpanelF).
AcomparisonofLipoD293(Ver.II)andLipofecatmineLTX(LTX)ongenerationofLentivirus(LV).ThreeCDNAswereco-transfectedwithLipoD293(Ver.II)andLipofectamineLTX(LTX)into293FTcells.AGFPvector,pHR-SIN-cppt-CMVEWP,wasusedtodeterminetiterofLV.1x105293Fcellsperwellwasplatedintoa24wellplatefollowedbyadditionofdifferentofamountsofthevectorsupernatant,1microliter(upperpanel)and10microliters(lowerpanel)respectively.5dayslater,thecellswaspassedwithFACS.Thenumbersattheupperrightcornerindicatethepercentageoftransducedcells.ThetitersofLVgeneratedwithLipoD293andL2Kwerequantifiedtobe8x10^6and3x10^6tu/mlrespectively
TechnicalInformation&Datasheet
-LipoD293(Ver.II)ReagentGeneralProtocol&DataSheet
-LipoD293(Ver.II)ReagentShortProtocol
-LipoD293(Ver.II)ReagentProtocolforTransfectingSuspension293andCHOCells
-LipoD293(Ver.II)ReagentforLentivirusProduction
-LipoD293(Ver.II)ReagentforrAAVProduction
-TechnicalNote&TransfectionTips
Torequestafreetrialsample,pleaseCreateAnAccountwithustoenteryourshippingaddressandemailusatorder@Signagen.com
Testimonials
IamabsolutelythrilledwiththeLipoD293transfectionreagent!IcomparedLipoD293toLipofectaminefortransfectionofplasmidstogenerateviralpseudoparticlesandWOW!Irecovered4timesmorevirususingtheLipoD293reagentthanLipofectamine!!ThisiswonderfulnewsformeandmymentorasitmeansthatIdon"thavetospendsomuchtime(andresources)ontransfecting293Tcellstorecovervirusforinvitroexperiments.MymentorwasalsoveryhappythatLipoD293costsmuchlessthanLipofectamine!!Wehavealreadyordered5mlsofLipoD293andIhavenowconvincedmylabmatestoswitchtoLipoD293sinceIgotsuchgoodresultswithit.Thanksformakingsuchagreatproduct!
--------ChristyLavine,Ph.D.,HarvardUniversity
IwantedtoupdateyouonthefreesampleofLipoD293thatyousenttous.IrecentlytransfectedsomePlat-E"sside-by-sidewithLipofectamine2000,andyourproductout-performedit!!!WearealsovalidatingitwithHeLacells,butotherwiseweareVERYhappywithwhatwehaveseenthusfar!Thankyouforsendingusthefreesample,itdefinatelymadetheSALEforus!
--------CatherineGallo,Ph.D.,UniversityofCincinnati
wehavenowtriedtheLipoD293DNAInVitrotransfectionreagenton293FTcells.InthefirstflaskweusedtheprotocolbyInvitrogenprovidedintheVirapowerboxinsert(Virapower,pBABE-GFPplasmid,plusLipofectamine).InthesecondflaskweusedLipoD293(30ul/6mlmedia)withthesameamountsofVirapowerandplasmidasinthefirstflask.Theresultswerestunning:LipoD293gavetwiceasmanytransformantsasLipofectamine2000...
--------ABetaTesterfromWayneStateUniversity
