Band on Western Blot are smeared - (Mar/25/2014 )
I am currently extracting nuclear proteins using a high salt buffer (400mM) and sonication. The band for Histone H3 (17kDa) is smeared and making it hard to quantify. I loaded 30ug protein/lane and ran on a 4-20% Biorad TGX gel. I have attached an image for reference. Any suggestions of how to fix the issue?
Try loading less protein, also try dialyzing the salt out of your samples.