western blot question
(by R.Jayakumar from roswellpark.org)
Thu Sep 27 13:49:29 EST 2007
A western blot or an SDS PAGE gel (stained)???? Generally if you only
need to see the bands, you can just stain the gel. If you are going to
do a blot, it is naturally assumed that youwish to do an immunoassay
with a antibody.
In an SDS-PAGE, you would get the heavy and light chains around the
50-60kDa and around the 25-30 kDa positions on a gel. I have observed
doubles at the both positions with the doublet at the 25-30 kDa better
separated than the higher molecular weight bands. Variation in
separation of the doublets (sometimes, I have also observed triplets)
varies with different antibodies. Does that answer your question?
"Rebecca Pickin" <RPickin from cvm.msstate.edu> wrote:
>I was wondering if anyone can help me. I am looking for a breakdown of
>what bands should appear on a western blot when you load the primary
>antibody onto the gel. I understand that to a certain extent various
>antibodies will be slightly different.
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