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Methanol substitute for Western?

Jennifer L. Potter jras at post.its.mcw.edu
Fri Dec 1 18:03:43 EST 1995


Dear Frank,

Our lab does quantitative Western blotting and we were having problems
getting complete transfer of our proteins.

We simply eliminated the MeOH from our buffer and have had great 
transfer ever since (43.2 g Glycine, 8.92 g Tris Base, 4 L DW).  

In Western blotting, many competing things are happening (i.e. SDS on the
proteins is what makes them move onto the nitrocellulose but SDS also
decreases the binding affinity of the nitrocellulose)

It's probably best to work out conditions empirically.

So I guess I don't know the answer to your question if EtOH is an okay
substitute for MeOH, but you may not need to use it at all.

Hope this helps,

Jennifer Potter
Grad Student
Medical College of WI (no,not Madison!)
  




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