I presume you've included as much as 0.01% SDS in the transfer buffer?
Another option is to use a semi-dry blotter. The two electrodes are closer
together and may get the protein out quicker. We have found that some
proteins that do not blot well in a liquid system do just fine in a semi-dry
blotter. Hopefully, you can borrow one.
Good Luck, Nancy Denslow.
At 05:34 PM 4/13/98 -0500, you wrote:
>A client has a 100kDa protein that does not electroblot from 7.5%
>acrylamide gels using CAPS/MeOH or Towbin/MeOH buffers. This is a selective
>phenomenon as all other proteins in his sample as well as ~100kDa standards
>are successfully blotted. We have reduced methanol concentration and
>included low concentrations of SDS without success. Longer blotting times
>are also of little avail.
>
>Any suggestions out there?
>
>Thanks,
>
>Larry Dangott
>
>
>
>
>
>Larry Dangott, Ph.D.
>Protein Chemistry Laboratory
>Texas A&M University
>College Station, TX 77843-3255
>(409) 845-2965
>
>
>
Nancy D. Denslow, Ph.D.
Scientific Director
Protein Chemistry Research Laboratory FEDEX address:
University of Florida ARB R3-234
PO Box 100245 Health Center 1600 Archer Road
Gainesville, FL. 32610 Gainesville, FL 32610
Phone:(352)392-9665
Fax: (352)846-1637
e-mail: Denslow@biotech.ufl.edu