Description
Basedonourproprietarypolymersynthesistechnology,PolyJetDNAInVitroTransfectionReagentisformulatedasabiodegradablepolymerbasedDNAtransfectionreagentthatensureseffectiveandreproducIBLetransfectiononHEK293,COS-7,NIH-3T3,HeLa,CHOandabroadrangesofhard-to-transfectmammaliancells.PolyJetreagentisabletoimmobilizeDNAmigrationduringelectrophoresisatverylowconcentrationandformtransfectioncomplexwithin5minutesatRT.Aremarkablefeatureofthereagentistherapidandcompletedegradationofpolymeraftertransfectioncomplexendocytosis(Figure1),leADIngtomuchlesscytotoxicity.PolyJetreagent,1.0ml,issufficientfor~667transfectionsin24wellplatesor~333transfectionsin6wellplates,providingaveryaffordablealternativetotheleadingproductsfortransfectingavarietyofcommonlyusedandhard-to-transfectmammaliancells.
Figure1.ACartoonShowingBiodegradationofPolyJetDNATransfectionReagentAfterEndocytosisofTransfectionComplex
Features
-Bio-degradableafterendocytosis
-Exceptionalhightitersofvirusproduction
-EquallygoodforverylongDNAs(>89kb)
-EquallygoodforbothsingleDNAtransfectionandmultiDNAco-transfection
-Highlevelsofrecombinantproteinproduction
-Simple&robusttransfectionprocedure
-Veryaffordable
StorageCondition
Storeat4°C.Ifstoredproperly,theproductisstablefor12monthsorlonger.
BroadTransfectionSpectrumforMammalianCellTypes
CellLines | Efficiency(%GFP) | CellLines | Efficiency(%GFP) |
| McArdle7777 Hep3D SHEP 3T3-442A COS-7 CV-1 D407 DHDPro.b 3LL B16-F10 BAEC BHK-21 CaSki CaCo2 CHO HCS-2/8 HEK-293 HeLa HLMEC H-MVEC Huh-7D12 ATT20 SK-N-SH C2C12 HepG2 | 65-70% | hESCs | 70% |
ExamplesShowingTransfectionEfficiencyofPolyJetDNAInVitroTransfectionReagentonCommonlyUsedCells
TransfectionefficiencycomparisonofPolyJetvs.lipofectaminePlusonChineseHamsterOvary(CHO)cells.HAtaggedbeta-tubulinCDNAwasdeliveredintoCHOcellswithPolyJet(leftpanel)andlipofectaminePlus(rightpanel)respectively.FITCconjugatedantibodyagainstHAtagwasutilizedtopickupHA-beta-tubulin(Green)whileaDM1aantibodywasusedtodetectendogenousalpha-tubulinfollowedbyprobingwithrhodamineconjugatedsecondaryantibody(Red).TheabovepicturewasprovidedbyDr.ShangYinofUniversityofTexasatHoustonMedicalSchoolascourtesy
AcomparisonshowingtransfectionefficiencyofPolyJetreagentvs.aleadingproduct,Lipofectamine2000onHEK293FTcells.HEK-293FTcellsweretransfectedwithGFPvector(pEGFP-N3)byPolyJet(leftpanel)andLipofectamine2000(rightpanel)respectively.ThecellswerevisualizedbyNikonEclipseFluorescencemicroscope24hoursposttransfection
AcomparisonshowingtransfectionefficiencyofPolyJetreagentvs.aleadingproduct,Lipofectamine2000onHepG2cells.HepG2cellsweretransfectedwithGFPvector(pEGFP-N3)byPolyJet(leftpanel)andLipofectamine2000(rightpanel)respectively.ThecellswerevisualizedbyNikonEclipseFluorescencemicroscope24hoursposttransfection
TransfectionefficiencycomparisonofPolyJetvs.FugeneHDonMDCKcells.AplainGFPDNAwastransducedintoMDCKcellswithPolyJet(leftpanel)andFugeneHD(rightpanel)reagentsrespectivelypermanufacturers"protocols.GFPandDAPIstainingwerevisualizedunderfluorescencemicroscopy48hoursposttansfection.TheabovecomparisondataandpictureswerecompletedandprovidedbyDr.GeZhouofNYUMedicalCenterascourtesy
AcomparisonshowingtransfectionefficiencyofPolyJetreagentvs.aleadingproduct,Lipofectamine2000onMDCKcells.MDCKcellsarenotoriouslyhardtotransfect.Withproprietary"ShavedCellTransfection"protocol,PolyJet(leftpanel)givesupto70%GFPpositivecellsvs.Lipofectamine2000(rightpanel)around5%efficiency.MDCKcellsweretransfectedwithGFPvector(pEGFP-N3)byPolyJet(leftpanel)andLipofectamine2000(rightpanel)respectively.ThecellswerevisualizedbyNikonEclipseFluorescencemicroscope36hoursposttransfection
AcomparisonshowingtransfectionefficiencyofPolyJetreagentvs.aleadingproduct,FugeneHDonLNCapcells.LNCapcellsweretransfectedwithGFPvector(pEGFP-N3)byPolyJet(leftpanel)andFugeneHD(rightpanel)respectively.ThecellswerevisualizedbyNikonEclipseFluorescencemicroscope24hoursposttransfection
AimageshowingexceptionaltransfectionefficiencyofPolyJetreagentonHumanembryonicstemcells (hESCs). ThehESCsgrowninE8mediumonGeltrexvs(leftpanel,DICimaging)wastransfectedwithpEF1α-GFP.TheGFPexpression(rightpanel)wasvisualizedbyNikonEclipseFluorescencemicroscope24hoursposttransfection. TheabovepictureswereprovidedbyDr.MarinaPryzhkovaofJohnsHopkinsUniversityascourtesy
Neuro2AcellstransfectedwithpEGFP-C1plasmidusingPolyJetInVitroDNATransfectionReagent.TheNeuro2AcellswerevisualizedbyNikonEclipseFluorescencemicroscopewithDICphaseimaging(left)andFITCimaging(right)24hourspost-transfection
ComparisonofcytotoxicityofPolyJetDNAInVitroTransfectionReagentwithL2Konprimarymurineskinfibroblast.Theprimarymurinefibroblastwasincubatedwiththeindicatedtransfectionreagents/pEGFP-C1(DNA)complexesabovefor4hoursinserum-freeDMEMHighGlucosemediumfollowedbyreplacementofcompleteserum-containingmedium.ThecellswerevisualizedbyNikonEclipseFluorescencemicroscopewithDICphaseimaging24hoursposttransfection
DataSheetandProtocols
-GeneralProtocolforTransfectingMammalianCells
-AShortProtocolforTransfectingMammalianCells
-AdvancedProtocolforTransfectingHard-to-TransfectMammalianCells
-AProtocolforTransfectingSUSPension293andCHOCells
-ProtocolforLentivirusProduction
-ProtocolforrAAVProduction
- TechnicalNote&TransfectionTips
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Testimonials:
Sorryforsooobigdelay,butnowIamtotallyinlovewithPolyJet.IhaveusedPolyJetforawhileonmouseESCswithrelativelygoodefficiency50~70%.
--------MarinaPryzhkova,Ph.D.,JHU
PolyJetTransfectionReagentworkedequallyaswellaslipofectamine2000,withlittleevidenceofcelldeathon293,PC-3and22RV1cells.Iwilldefiantlyconsiderswitchingover.
-----TiffanyWallace,Ph.D.,NCI/NIH
Ionlydidsidebysidewiththetestingsample(PolyJet)andLipo2000withGFPtransfectiononCOS-7cells.Theresultwasverygood.PolyJetwasevenbetterthanL2K.
------FengQiao,Ph.D.,NEI/NIH
ItestedthesampleofPolyJetonmyNIH-3T3mousefibroblaststhisweekend.TheresultsweremuchbetterthanLipofecatmineLTX.I"mattachingapowerpointslidewithmyresults(Ididnotquantifythe%transfectionefficiency,butthepicturesgetthepointacross).Ifoundthattheprotocolfordifficult-to-transfectcelllinesworkedbetterthanthestandardprotocol.
-----StephanieMurphy,Ph.D.,DartmouthCollege
Ihadchancetotryyourproductfinally.Itwasgreatsuccess.IusedHeLacellsandgot10%transfectionefficiency(<0.1%forLipofectamine).Thankyou!IwaswonderingifIalsotryGenJetPlusDNAInVitroTransfectionReagent?Accordingtoyourwebsite,thereagentworksbetterthanregularPolyJet.
-------YumiUetake,Ph.D.,UMASS
ItestedPolyJetanditlooksgreatonMDCK.Weplacedorder.Thankyou!
-------GeZhou,Ph.D.,NYU
Wearehappytoprovidefeedback.PolyJetworkedverywellforusinHepG2cells,wegotapproximately80%efficiencywithpMAXGFPplasmid,byfollowingtheconditionsinyoursuggestedprotocol.WeranacomparisonwithLipofectamine,whichonlyshowedapproximately20-30%transfectionefficiency.Weareplanningexperimentsandwillbeorderingmoresoon.
-------EmilyMcallister,PBRC
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