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about western blot analysis - (May/03/2013 )

I have done western blot analysis with tissue proteins. When I visualized it with chemiluminescent gel imager all the bands came as columns. I repeated it 3 times and i got the same results. So I wanted to know what is the reason for getting that kind of columns instead of bands. Please reply.

Furthermore I have used the SDS-PAGE gel and follow the instructions according to the manual.

Thank u.


The options are quite a few - protein content, protein loading, poor running, poor transfer, poor denaturation, antibody specificity (primary and secondary), over exposure, insufficient washing, stringency conditions, blocking conditions....