western blot question

Jayakumar, R via methods%40net.bio.net (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?

-----Original Message-----

"Rebecca Pickin" <RPickin from cvm.msstate.edu> wrote:
>Hi everybody,
>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.

This email message may contain legally privileged and/or confidential information.  If you are not the intended recipient(s), or the employee or agent responsible for the delivery of this message to the intended recipient(s), you are hereby notified that any disclosure, copying, distribution, or use of this email message is prohibited.  If you have received this message in error, please notify the sender immediately by e-mail and delete this email message from your computer. Thank you.

More information about the Methods mailing list