Western Blot problems

Andreas Kampkötter andreas.kampkoetter at uni-duesseldorf.de
Thu Aug 20 10:05:04 EST 1998


Is there somebody, who can provide a reliable recepy to get
membrane-associated proteins into solution for Western Blot analysis?
The protein, I´m working on, is about 25 kD in size and farnesylated. For
this reason I assume that it is a membran-associated protein. In Western
Blot experiments I can sometimes detect a band of the expected size using an
affinity-purified antibody and a heterologous  commercial available
antibody. But in most experiments I detect only bands of a high molecular
weight and no band at 25 kD. I ´ve homogenized the worms, we are working
with (schistosomes; plathyhelminthes), in several different lysis buffers
(RIPA, SDS, Urea, GuHCl etc.) with and without sonification.
I hope that somebody can help me with my problems.
Thanks,
     Andreas



 
Andreas Kampkoetter
Institute for Genetics
Heinrich Heine Universitaet
Universitaetsstr. 1
D-40225 Duesseldorf/Germany

Tel.:   (49)-211-811-2077
e-mail: andreas.kampkoetter at uni-duesseldorf.de




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