🚀 go-pugleaf

We have had similar problems staining western blots of plant proteins.  We
use biotinylated marker proteins, and at first we'd incubate the
nitrocellulose membrane + proteins with the primary antibody, then with a
combination of HRP-labelled secondary plus avidin-HRP.  This produced lots
of bands all over the place.  Now, we cut off the lane with the markers and
incubate it separately with avidin-HRP, and blot the rest with primary then
HRP-secondary.  None of the extra bands show up if we do this.  So the
avidin is binding to something.  Of course, you can't do this with your
tissue!  Perhaps an alkaline phosphatase system would be the way to go?


NOTE:  NEW PHONE AND FAX NUMBERS AS OF 8 MAY 1995
____________________________________________________________
   Rosemary White                       __    /
   Department of Ecology              _/  \__/ \
      and Evolutionary Biology       /          \
   Monash University                /  Australia \
   Clayton, Victoria 3168           \   ____     /
   phone  61-3-9905 5670             \_/    \_*_/
   fax    61-3-9905 5613                     __
   email  r.g.white@sci.monash.edu.au        \/