protein quantification

Dr Engelbert Buxbaum engelbert_buxbaum at
Wed Feb 15 07:51:52 EST 2006

Rohatgi, Tanuja wrote:

> I would like to use the conditioned medium for the
> experiments but do not know how to quantify the amount of my protein of
> interest in this conditioned medium. I do have a specific monoclonal Ab
> against the protein and it works well on the blot. Can you suggest to me
> how can I quantify the protein by Coommassie or some other means.

Dye binding is used to measure total protein. To determine the
concentration of a specific protein you can use an ELISA, dot blot or
spot blot assay, since you have a specific antibody. Western blotting is
at best semi-quantitative. In any case you will also need a purified
sample of your protein in order to calibrate the assay.

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