IUBio

[Protein-analysis] IEF transfer

Brandon Hostetler hostetle at hotmail.com
Wed May 10 08:59:16 EST 2006


I saw your comment on an IEF transfer problem, and I was hoping you could 
help me.
First, any help you can offer would be appreciated, but if you are unable, 
thank you anyway.
To my point, I am trying to do an IEF slab gel and I have tried both 
nondenaturing and denaturing conditions in a pH range of 3-7.  After I do my 
transfer I silver stain the gel and I have found a great deal of proteins 
still on the gel.  I have tried to transfer in Acetic Acid I have tried 
transfering with 192mM Glycine, 50mM Tris, 0,1%SDS, 20%Methanol, but still 
not all the proteins are transfering.  The only proteins that show up in 
Ponceau are those near the cathodic end.  Do you have any suggestions?

Thanks

Brandon Hostetler

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