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Trans-Blot®SDSemi-DryElectrophoreticTransferCellInstructionManualCatalogNumber170-3940ForTechnicalServiceCallYourLocalBio-RadOfficeorintheU.S.Call1-800-4BIORAD(1-800-424-6723)NoteToinsurethebestperformancefromtheTrans-BlotSDsemi-dryelectrophoretictransfercell,becomefullyacquaintedwiththeseoperatinginstructionsbeforeusingthecelltotransfersamples.Bio-Radrecommendsthatyoufirstreadtheseinstructionscarefully.Thenassembleanddisassemblethecellcompletelywithouttransferringsample.Afterthesepreliminarysteps,youshouldbereadytotransferasample.Bio-RadalsorecommendsthatallTrans-BlotSDcellcomponentsandaccessoriesbecleanedwithasuitablelaboratorycleaner(suchasBio-RadCleaningConcentrate,catalognumber161-0722)andrinsedthoroughlywithdistilledwater,beforeuse.WarrantyBio-RadLaboratorieswarrantstheTrans-BlotSDsemi-dryelectrophoretictransfercellagainstdefectsinmaterialsandworkmanshipfor1year.Ifanydefectsoccurintheinstrumentduringthiswarrantyperiod,Bio-RadLaboratorieswillrepairorreplacethedefectivepartsfree.Thefollowingdefects,however,arespecificallyexcluded:1.Defectscausedbyimproperoperation.2.RepairormodificationdonebyanyoneotherthanBio-RadLaboratoriesoranauthorizedagent.3.UseoffittingsorothersparepartssuppliedbyanyoneotherthanBio-RadLaboratories.4.Damagecausedbyaccidentormisuse.5.Damagecausedbydisaster.6.Corrosionduetouseofimpropersolventorsample.Thiswarrantydoesnotapplytopartslistedbelow:1.Platinumplateelectrode.Foranyinquiryorrequestforrepairservice,contactBio-RadLaboratoriesaftercon-firmingthemodelandserialnumberofyourinstrument.Model___________________________________CatalogNumber__________________________DateofDelivery___________________________WarrantyPeriod__________________________SerialNumber____________________________InvoiceNumber___________________________PurchaseOrderNumber____________________TableofContentsPageSection1Introduction..................................................................................................11.1Specifications.............................................................................................................1Section2EquipmentandReagents............................................................................22.1EquipmentandAccessories.......................................................................................22.2RelatedInstruments....................................................................................................42.3ChemicalReagents.....................................................................................................4Section3SafetyInstructions.......................................................................................5Section4Trans-BlotSDAssembly.............................................................................64.1PreparationforBlotting.............................................................................................64.2AssemblyoftheUnitforStandardTransfers............................................................74.3AssemblyoftheUnitforAcidicTransfers...............................................................10Section5BufferFormulation......................................................................................10Section6ExamplesofSpecificProtocols...................................................................116.1SDS-ProteinBlotting.................................................................................................116.2DNABlotting(ForacrylamidegelswithDNA250bpto~1kb)............................126.3DNA&RNABlotting(ForagarosegelswithDNAupto23kb,RNAupto3.5kb)......................................................................................................12Section7PropertiesofProteinBlottingMedia.........................................................12Section8TroubleshootingGuide................................................................................138.1PoorTransfer..............................................................................................................138.2PoorBindingtoNitrocelluloseMembrane................................................................148.3HighBackgroundAfterIncubationwithAntibodyProbes;NonspecificorNonquantitativeDetection.....................................................................................148.4PoorDetectionSensitivityorNoReactivity.............................................................15Section9References.....................................................................................................15Section1IntroductionBlottingwasfirstperformedbySouthern1in1975withthetransferofDNAfromagarosegelstonitrocellulosemembranes.BlottinghassubsequentlybeenappliedtoRNA2-4andprotein5,6frombothagaroseandpolyacrylamidegels.MembranematerialshavebeenexpandedtoincludePVDFforimprovedproteinbindingcapacity.Toovercometheinefficiencyofcapillarytransfers,electriccurrenthasbeenadoptedforelutingproteinsfrompolyacrylamidegels,asfirstdescribedbyTowbinetal.7in1979.Sincethattime,electrophoretictransferhasal