Loading control for western blot of serum samples...

1D, 2D, HPLC, SDS PAGE, etc. Talk about it here.
Albumin Member
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Loading control for western blot of serum samples...

Postby Biomarker » Mon Dec 10, 2012 2:25 am


Can anybody please comment on the use of loading control? I am working on glycation PTM (non-enzymatic process). I want to ensure whether i have loaded equal protein on gel or not.

How about the use of commassie staining as loading control.


Angiotensin Member
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Western of a housekeeping protein is better for a loading control

Postby Jen_G » Wed Feb 20, 2013 10:38 am

Once a gel is stained with Coomassie you can do a western blot on it. For a rough estimate you can ponceau stain the blot but using another primary antibody for a protein that doesn't change is better. That way you can quantitate the bands to prove that you have equally loaded your lanes. Popular ones are actin, beta-tubulin, or AKT.

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Postby MassSpecMan » Wed Feb 20, 2013 12:23 pm

If you are using the yeast model system, our lab has used PGK1 in the past as a loading control. It is a very good antibody at 1:1000 dilution, and gives very little background.

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