MeOH in Western Buffer

Malcolm Moos Jr. moos at helix.nih.gov
Sun Mar 12 18:27:08 EST 1995


lhom at OCF.Berkeley.EDU (Louis Hom) wrote:
>
>The original rationale for methanol was to enhance interactions of 
proteins with nitrocellulose (especially smaller, highly charged ones)
and to prevent swelling. Most of the time, neither consideration is 
serious. With large, high aspect ratio proteins (myosin), methanol can
drastically reduce transfer efficiency. I never use it. The 10 mM CAPS
buffer suggested by Matsudaira for protein sequencing has never failed
in my hands, is very inexpensive, and runs at low amperage and therefore
temperature. The only thing you might need to do on occasion is to add
very small amounts of SDS (around 0.005%) if you have a protein that 
won't transfer.



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