SDS-PAGE for purity
dk at no.email.thankstospam.net
Tue Jul 16 18:14:14 EST 2002
drabena at hotmail.com wrote:
>I am currently running SDS-PAGE's to calculate the "percent area" of a single
> protein. Can anyone tell me how to run this assay to determine the percent
There is no truly good way to run such an assay. Conventional
scanning/integration of total lane gives absolutely wrong results.
The best way I know is to i) determine total protein concentration
in your sample, ii) load in parallel a series of known amounts
of [some standard] protein, iii) compare intensities, deduce amount
of the given band in your sample and thus deduce % of total.
With some experience, simple eyeballing estimates come as
close to reality as any "quantitative" method (that is, still
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