Western blot problem

Ramaswamy K. Iyer riyer at pathology.medsch.ucla.edu
Tue Jan 20 01:32:33 EST 1998

Help! I am getting unusual signal on my western blots using the Amersham
ECL kit. I am probing for B-actin but as well as getting a band at the
correct size, I also see a smear of signal at the outside edges of each
lane stretching from the top to the bottom of the blot. I am using a 6%
separating gel and a 4% stacking gel and standard buffer system. The
blotting is done overnight at 4 degrees and the blot is blocked with 10%
milk in PBST. The primary and secondary antibodies have been used before
at these same dilutions and conditions without these problems. I am
loading 25 ug of lysate. Thanks for any suggestions that you may have.

Ram Iyer
riyer at ucla.edu

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