Western blot-polysomes problem

JMORELLI at MAINE.MAINE.EDU JMORELLI at MAINE.MAINE.EDU
Sat Nov 5 02:48:01 EST 1994


I am having difficulty doing a western blot for ef-1a which associates
with polyribosomes.  The Ab works fine against total plant extract,
but when blotting th polysome component, every band lights up.
I am using the Amersham chemiluminesence protocol, blotting
to nitrocellulose, and blocking with BSA.  I tried nonfat dry milk
and dextran sulfate, higher molarity NaCl, higher tween, more dilute
Ab 1st and second, reduced the polysome protein concentration to 5 ug
instead of 20.  None of these things helped. Any ideas?
Thanks in advance,
Jim Morelli
University of Maine
207-581-2828
jmorelli at maine.maine.edu



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