western blotting

[davidmc at scripps.edu]

   We have been having difficulty with detection of a proteolytic fragment
of a protein by immunoblotting.  The problem seems to be that the fragment
is easily washed off PVDF membranes.  This may be due to unusual
hydrophobic/philic properties of the protein in question.  We have tried
modifications of transfer protocols and other membranes but have not seen
any improvement.  Has anyone had a similar experience or had any
experience with cross linking proteins to blot membranes?

   Please respond by e-mail to davidmc at scripps.edu