JournalofVisualizedExperimentswww.jove.comCopyright©2011JournalofVisualizedExperimentsPage1of2MaterialsListfor:TransmembraneDomainOligomerizationPropensitydeterminedbyToxRAssayCatherineJoce1,AlyssaWiener1,HangYin11DepartmentofChemistryandBiochemistry,UniversityofColoradoatBoulderCorrespondenceto:HangYinathang.yin@colorado.eduURL:http://www.jove.com/video/2721DOI:doi:10.3791/2721MaterialsNameCompanyCatalogNumberCommentsBamHIrestrictionenzymeInvitrogen15201023InvitrogenenzymeswerefoundtobemoreefficientthanalternativesuppliersNheIrestrictionenzymeInvitrogen15444011Invitrogenenzymeswerefoundtobemoreefficientthanalternativesuppliers15mLculturetubesFisherScientific14-956-1JSOCmediaTEKnova,Inc.S0225Madeuptotheappropriatevolumeandsterilizedbyautoclaving.LBmediaSigma-AldrichL7275Madeuptotheappropriatevolumeandsterilizedbyautoclaving.ChloramphenicolSigma-AldrichCO378Stocksolutionof30mg/mLinethanolstoredinfreezerArabinoseFluka10839Stocksolutionof2.5%(w/v)inwaterstoredinfreezerNa2HPO4Sigma-AldrichS9390NaH2PO4Sigma-AldrichS9638KClMallinckrodtBakerInc.6858-06MgSO4.7H2OSigma-Aldrich63138Sodiumdodecylsulfate(SDS)Sigma-AldrichL60262-Nitrophenylβ-D-galactopyranoside(ONPG)Sigma-Aldrich73660Z-buffer16.1gNa2HPO45.5gNaH2PO40.75gKCl0.246gMgSO4Makeupto1l,pH7.0Z-buffer/chloroform200mLβ-mercapt–thanol,2mLchloroform,makeupto20mLwithZ-buffer.Vortexfor1min,JournalofVisualizedExperimentswww.jove.comCopyright©2011JournalofVisualizedExperimentsPage2of2centrifugefor1minat800rpm.Makefreshforeachplate.Z-buffer/SDS160mgSDSdissolvedin10mLZ-bufferZ-buffer/ONPG40mgONPGin10mLZ-buffer.Makefreshforeachplateβ-mercapt–thanolCalbiochem444203Anti-MBPmonoclonalantibody(HRPconjugated)NewEnglandBiolabsE8038SMinimalmediawithmaltose1xM9salts,0.4%maltose,1mg/mLthiamin,2mMMgSO496-wellflatbottomplateSarstedtLtd83.1835.300Plate-readerBeckmanCoulterInc.DTX880MultimodeDetectorWaterbathVWRinternational89032-204ShakingincubatorFormaScientific
