Neg control fopr western? - (Jan/28/2003 )
What are the best or most used negative controls for western blot. Is it possible to use serum or IGg from the same species the primary antibody is made in?
we use only the secondary ab for negative control, so you can see, whether it reacts directely with the blot.
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Usually we stain the same sample with IgG from the species the antibody is raised in to
differentiate between nonspecific and specific bands, like rabbit anti-X, would use Rabbit IgG as
negative control. It would be good if you also have the positive control mentioned on
each antibody sheet if you have the cell line with the protein/protein sample in your lab.
The specific band should only occur in the specific antibody lane, and should be of
strong intensity.(modify antibody dilution to get the best result.)