MeOH in Western Buffer

John Fox J.R.Fox at shu.ac.uk
Tue Mar 14 10:10:37 EST 1995


In article <3jq49i$iq2 at agate.berkeley.edu>, lhom at OCF.Berkeley.EDU (Louis Hom) says:
>
>My labmates are invariably astonished to hear that I don't put methanol in
>my Western transfer buffer.  But I never have, and I still get nice transfer.
>Why do people put MeOH in their buffer? Does its absence not affect me because
>of the membrane I use (nitrocellulose)? 
>-- 
>______________________________________________________________________________
>Lou Hom >K'93           "It has been suggested that...gene splicing and cloning 
>lhom at ocf.berkeley.edu    are so simple that one can successfully carry them out 
>                         in [any] modern kitchen. In spite of my considerable
> Quote by                respect for the potential of food processors and radar
>     B.R. Levin          ranges, I very much doubt this to be the case."
>------------------------------------------------------------------------------------------------------------------                        
There are two reasons why methanol is included in the Western transfer buffer :

A) Methanol can in some cases restrict elution of protein from the gel.  

B) Nitrocellulose requires 20% methanol in the buffer for optimal protein binding. 

Elimination of methanol results in increased transfer efficiency but does diminish
protein binding to nitrocellulose.


Regards

John 



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