产品资料

Polyplus Transfection/jetPEI^®-HUVEC/Polyplus Transfection-163152/1 Ea

Efficienttransfectionofendothelialcells(upto50%inHUVEC)
Asefficientaselectroporation
Easy-to-useprotocol
GoodcellviABIlity
CompatIBLewithserumandantibiotics

Specifications:

Reagent	jetPEI^®-HUVEC
Moleculedelivered	DNA
Applications	DNAtransfectionofendothelialcells
Celltypes	Endothelialcells
Numberoftransfections	0.5mlofjetPEI^®-HUVECtransfectionreagentissufficienttoperformupto125transfectionsin24-wellplatesor40transfectionsin60-mmplates
Storage	4°C,forupto12months
Providedwith	150mMNaCl

Summary:

jetPEI^®-HUVECisapowerfultransfectionreagentoptimizedforthetransfectionofprimaryhumanendothelialcellssuchasHUVEC(Humanumbilicalveinendothelialcells).Transfectionefficienciesupto50%havebeenreachedwiththisreagent.

Orderinginformation:

|||108-05N##0.5ml##50ml##|||

Description:

jetPEI^®-HUVECleadstoimprovedtransfectionefficiencyofendothelialcells

ComparedtojetPEI^®,ourstandardandversatiletransfectionreagent,jetPEI^®-HUVECprovidessignificantlyhighertransfectionefficienciesinHUVECcells(Fig.1).

jetPEI-HUVEC - Comparison jetPEI — **Fig.1:TransfectionefficiencyandtoxicityofjetPEI^®andjetPEI^®-HUVECinHUVEC.**Cellswereincubatedfor4hwithtransfectioncomplexesin2%serummedium.Luciferaseactivitywasmeasured24haftertransfection.

Indeed,jetPEI^®-HUVECinduceslesscellulartoxicitythanjetPEI^®.UsingtheoptimalconditionsforjetPEI^®-HUVEC,transfectionefficienciesof50%havebeenreachedasobservedwithafluorescentproteinreportergene(Fig.2).

jetPEI-HUVEC - FACS — **Fig.2:FACSanalysisofHUVECcellstransfectedwithaplasmidencodingtheEGFPfluorescentproteinusingjetPEI^®-HUVEC.**Transfectionwasperformedin24-wellplatewith2µgofplasmidand4µlofjetPEI^®-HUVEC(N/P=5)andin2%serum.Complexeswereincubatedfor4hwiththecells.FACSanalysistoquantifyEGFPexpressionwasperformed24haftertransfection.

Optimizationoftheexperimentalconditionsforhightransfectionefficiency

Theeffectofserumconcentrationandincubationtimeontransfectionefficiencywasanalyzed(Fig.3).

jetPEI-HUVEC - Incubation with serum — **Fig.3:Effectoflow(2%)versushigh(30%)serumconcentrationsontransfectionefficiencyofjetPEI^®-HUVEC**.HUVECcellsweretransfectedfor2,4or24hwith2µgofpCMVLucplasmidDNAand4µlofjetPEI^®-HUVECin24-wellplates.Luciferaseactivitywasmeasured24haftertransfection.

Transfectionexperimentsperformedinlow(2%)orhigh(30%)concentrationsofserumhaveshownthatjetPEI^®-HUVECismostefficientwhenthecellsaregrowninlowserumconcentrations.GeneexpressionreachesamaximumforHUVECafter4hincubationwithDNA/jetPEI^®-HUVECcomplexes.

jetPEI^®-HUVECcomparesparticularlyfavorablytoothernon-viraltransfectionreagentswhethercationiclipidsorpolyamine(Fig.4).

jetPEI-HUVEC - comparison transfection efficiency — **Figure4:ComparisonoftransfectionefficiencyofjetPEI^®-HUVECandothercommerciallyavailablereagents(P1-P4**).Cellsweretransfectedusingoptimalconditionsestablishedforeachreagent.

Easytouseprotocol

ThejetPEI^®-HUVECprotocolisassimpleasthejetPEI^®one:MixtheDNAwiththereagenttoformcomplexesandsimplyaddthemixturetothecells.jetPEI^®-HUVECiscompatiblewithserumandantibiotics,thuseliminatingtheneedformediachange.Proteinexpressionisdetermined24hto72hpost-transfection.

Applications:

jetPEI^®-HUVECisperfectlysuitedforplasmiddelivery(DNA,shRNAormiRNAplasmids)tofragilecells,suchasprimaryendothelialcells.

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Citations:

HereisaselectionofrelevantreferencesusingjetPEI-HUVEC,moreareavailableinourPolyplus-transfectionDatabase.Feng,J.,Zhang,Y.,Xing,D.(2012).Low-powerlaserirrADIation(LPLI)promotesVEGFexpressionandvascularendothelialcellproliferationthroughtheactivationofERK/Sp1pathway.,CellSignal 24,1116.Ganguly,A.,Yang,H.,Zhang,H.,Cabral,F.,Patel,K.D.(2013).Microtubuledynamicscontroltailretractioninmigratingvascularendothelialcells.,MolCancerTher 12,2837.Jin,H.G.,Yamashita,H.,Nagano,Y.,Fukuba,H.,Hiji,M.,Ohtsuki,T.,Takahashi,T.,Kohriyama,T.,Kaibuchi,K.,Matsumoto,M.(2006).Hypoxia-inducedupregulationofendothelialsmallGproteinRhoAandRho-kinase/ROCK2inhibitseNOSexpression.,NeurosciLett408,62-7.Wen,H.C.,Kao,C.,Hsu,R.C.,Huo,Y.N.,Ting,P.C.,Chen,L.C.,Hsu,S.P.,Juan,S.H.,Lee,W.S.(2013).Thy-1-inducedmigrationinhibitioninvascularendothelialcellsthroughreducingtheRhoAactivity.,PLoSONE 8,e6150.Cristofaro,B.,Stone,O.A.,Caporali,A.,Dawbarn,D.,Ieronimakis,N.,Reyes,M.,Madeddu,P.,Bates,D.O.,Emanueli,C.(2010).Neurotrophin-3isanovelangiogenicfactorcapableoftherapeuticneovascularizationinamousemodeloflimbischemia.,ArteriosclerThrombVascBiol30,1143.

Quality:

EverybatchofjetPEI^®-HUVECistestedinatransfectionassay.Typically,transfectionofafireflyluciferasegeneunderthecontroloftheCMVpromotergives10⁹RLU(relativelightunit)/mgofprotein.ThevalueforeachbatchisindicatedontheCertificateofAnalysis.

Testimonials:

« SofaramongalltheDNAtransfectionreagentstoHUVEC,yourjetPEI-HUVECisthebest.Thehighestefficiencywasabout30-50% »

HongZ,UniversityofCalgary,Canada

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Polyplus Transfection/jetPEI®-HUVEC/Polyplus Transfection-163152/1 Ea