Western Blot Problem

Ramaswamy K. Iyer riyer at pathology.medsch.ucla.edu
Tue Jan 20 21:41:55 EST 1998


Help! I am getting a bizarre signal on my western blots. I am using the
Amersham ECL detection system and the signal shows up not only as the
band I am trying to detect (B-actin) but also as vertical streaks in
between the lanes stretching from the top of the blot to the bottom. I
am using a 6% separating gel and a 4% stacking gel and I am using a
standard buffer system. I am using 10% milk in PBST to block overnight
at 4 degrees. The primary and secondary antibodies have been used before
at these concentrations and have given good results. Any advice would be
greatly appreciated.

Ram Iyer



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