JournalofVisualizedExperimentswww.jove.comCopyright©2012CreativeCommonsAttribution-NonCommercialLicensePage1of1MaterialsListfor:HighThroughputSequentialELISAforValidationofBiomarkersofAcuteGraft-Versus-HostDiseaseBryanFiema*1,AndrewC.Harris*1,AurelieGomez1,PraechompooPongtornpipat1,KellyLamiman1,MarkT.VanderLugt1,SophiePaczesny11PediatricBloodandMarrowTransplantProgram,UniversityofMichigan*TheseauthorscontributedequallyCorrespondenceto:SophiePaczesnyatsophiep@umich.eduURL:http://www.jove.com/video/4247DOI:doi:10.3791/4247MaterialsNameCompanyCatalogNumberCommentsHumanIL-2RalphaDuosetR&DSystemsDY223HumanHGFDuosetR&DSystemsDY294HumanIL-8OptEIAKITIIBectonDickinson550999Ab-MatchASSEMBLYHumanPAP1(REG3α)KitMBLInternational5323Ab-MatchUNIVERSALKitMBLInternational5310HumansTNFRI/TNFRSF1ADuosetR&DSystemsDY225HumanTrappin-2/ElafinDuosetR&DSystemsDY174796-wellpolystyreneconicalbottomplatesThermoScientific249570UsedforplasmasourceplatesCostarhalf-wellhigh-binding96-wellplatesCorning3690ForIL-2Rα,HGF,TNFR1andelafinELISAsNunc384-wellMaxiSorpplatesNunc464718ForREG3αElisaHyClonePhosphateBufferedSaline,1xThermoScientificSH30256.02BovineSerumAlbumin,FractionV,HeatShockTreatedFisherScientificBP1600-100BlockerBLOTTOinTBSThermoScientific37530BlockingagentforIL-2RαHGFandTNFR1ELISAsDulbecco'sPhosphate-BufferedSalineGibco21600-069WashbufferforIL-2Rα,HGF,ElafinandTNFR1ELISAsTMBPeroxideSusbtrateKirkegaardandPerryLaboratories50-76-00Tween20AcrosOrganics233362500WashbufferforIL-2Rα,HGF,ElafinandTNFR1ELISAsSulfuricAcidSigma-Aldrich84720(Dilutedto2N)forstopsolution
