Instruction Manual

3.7 Transfer Using Traditional Semi-Dry Consumables
TypicalprocedureutilizingtheTrans-BlotTurbosystemandconventionalsemi-drywesternblotting
consumablesisdetailedbelow:
1. EquilibratethegelinTowbintransferbuffer(25mMTris,192mMglycinepH8.3,20%MeOH)for10
min.
2. Soaktwopiecesofextra-thick(2.4mm)lterpaperintransferbuffer.Sixpiecesofthick(0.8mm)
lterpapercanbeusedifextra-thickpaperisnotavailable.
3. Whilethegelisequilibrating,prepareatransfermembrane.Wetanitrocellulosemembranebriey
intransferbufferorPVDFmembraneinmethanolorethanolfor30sec,thenwashinwaterfor1–2
min,andequilibrateintransferbufferforatleast10minwithagitation.
4. Assemblethetransfersandwichonthecassettebase(anode)byplacingonepieceofwetextra-
thickor3piecesofthicklterpaperonthebottom,thenthemembrane,thegel,andnally,
theremainderofthewetlterpaperontop.Usetheblotrollertoremoveairfrombetweenthe
assembledlayers(Figure19).
5. Oncethestacksarepositionedinthecassettebase,placethecassettelidonthebase.Thelidis
reversible,butensurethattheelectricalcontactstcloselyintotheslotsinthebase.Pressthelid
downrmlyandturnthedialclockwisetoengagethelidpinsintothelockingslots.
6. Loadasecondcassetteifdesired.RefertoTable1fortheappropriatecombinationsofgelsthat
canbecombinedinasinglerun.
7.  Slidethecassette(withthedialfacingup)intothebayuntilitmakescontactwiththemagnetic
interlockandyouhearaclick.Cassettescanbeinsertedintothebaysinanyorder,withorwithout
powertothesystem.
8. SelecttheLISTbuttonfromtheHomemenuandtheSTANDARD SDtransferprotocolfromthe
Bio-Radpreprogrammedprotocolsortheuser-denedprotocolofchoice.
9. Toinitiatetherun,pressthenavigationbuttonthatcorrespondstoA:RUNforthecassetteinthe
upperbayorB:RUNforthecassetteinthelowerbay.
Forfutherinformation,refertotheBio-RadProteinBlottingGuide:AGuidetoProteinDetectionand
Blotting,bulletin2895,fordetailedinformation.Bulletin2895canbedownloadedfromourwebsite,
www.bio-rad.com,asaPDFle,orcontactTechnicalSupportat1-800-424-6723.
Fig.19.Properassemblyofblottingsandwichusingtraditional
consumables.
Top(–)cassette
electrode(cathode)
Topionreservoir
stack
Blottingmembrane
Bottomionreservoir
stack
Bottom(+)cassette
electrode(anode)
Gel
TechSupport:1-800-424-6723•www.bio-rad.com15
Trans-Blot
®
Turbo™BlottingSystem