I am running a slab ief gel, using ph range 4-6,5. I am trying to transfer my ief seperated proteins to NC membrane but there is no result.
Before trasnfer i equilibrate my ief gel in : 192mM Glycine, 50mM Tris, 0,1%SDS, 20%Merhanol and i am using the same buffer for trasfering. Next im staining me Nc membrane using Ponceau, but there is no protein on membrane. Finally, i have no signal, using my monoclonal antibody and hyperoxidase.
Any suggestion? May my protein is missed during equilibration?
Thanks in advance for any idea
Hellenic Institute Pasteur
Departement of Biochemistry
GR 11521, Athens
pstasina at mail.pasteur.grpstasinar at mailhost.bio.auth.gr
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