IUBio

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



More information about the Proteins mailing list

Send comments to us at biosci-help [At] net.bio.net