BCA-Protein-Quantification: Time Drive?

Peter pxpst2 at SPAM.SUXS.unixs.cis.pitt.edu
Sun Aug 9 14:12:58 EST 1998


In article <35cab1d3.1944564 at news.fu-berlin.de>,
deleteme_cpeter at zedat.fu-berlin.de wrote:

> Hi all!
> 
> I use the bicinchoninic-acid-assay in a 96well format to
> determine/estimate protein concentrations. I always have a
> standard-dilution-row on every plate.
> 
> When I tried to determine the concentration of a simple yeast-extract,
> I noted the following:
> 
> - the slope of the calibration-line (mg as a function of A550)
> decreases with time (15min to 3hrs)
> 
> - the concentration of my extract also decreases with time!!!!
> (calculated with corresponding slope!)
> 
> As the assay is designed to be read after 30min and 30degrees C - when
> do I get the _correct_ concentration?????????????????
> 

As long as all decreases occur to all samples , It does not matter. 
Incubation of the reaction at 37 C for 30 minutes and then read should be
fine.  The key is that all are read at the same time( samples and
calibration curve)  variability will take care of itself.

Peter

-- 
"Don't you eat that yellow snow
            watch out where the Huskies go"    FZ

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