If using Unicorn, 1)Open your chromatogram in Evaluation window. 2)Go to Operations-> Pool 3)Choose which baseline estimation suits you, define the pools (numbered rulers that appear under the chromatogram) 4)type in the path length (2 or 10 mm for UV-900 and 2 mm for UPC-900) 5)type in the mass extinction coefficient (Molar extinction coefficient/Mw in daltons) 6) get your answer from the table at the bottom of the screen The reading will make sense for pure protein. Also note that for the most accurate result you need to know the exact path length which varies a little bit from cell to cell (or so they say).
Regards,
Dmitry
On 2014-01-15, at 10:09 AM, Karel Chaz wrote:
> Dear all,
>
> A question for the biochemistry-inclined folks in the bb; how do I calculate
> protein concentration of chromatography fractions, starting from Abs280 from
> the UV monitor? I know I could figure it out myself if I really tried, but
> why bother when I have access to so many brilliant minds....
>
>
> Thanks to all,
>
> K
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