Artifact by coomassie blue in amino acid analysis?
Achim Recktenwald, PhD
achim at ibex.ca
Mon Mar 3 08:01:01 EST 1997
Stephen P. Driska PhD wrote:
> newera at plaza.snu.ac.kr wrote:
> : Can coomassie blue cause an artifact of proline content in
> amino acid analysis
> : of a protein?
> : I have amino-acid-anayzed my protein 3 times, which is transfered on
> : PVDF membrane and stained with coomassie blue. Each time, I have got
> : a different result in terms of proline content but the contents of
> : the other amino acids have been measured almost the same.
> : What is the possible causes of this problem?
> : --
> : email : newera at plaza.snu.ac.kr
> : address :
> : Lee, Ji Hyun
> : Laboratory of Physical Pharmacy(Prof. Lee, Bong Jin)
> : Seoul National University
> : College of Pharmacy
> : Shinlim-Dong, Kwanak-Gu
> : Seoul 151-742, Korea.
> It's an obvious thing, but could you try removing the coomassie blue
> from the PVDF? Also, it might help people suggest an answer if you said what
> type of machine or column was being used for the analysis. A few years ago
> we were having a problem with coomassie blue and the Waters PicoTag system,
> but we had enough protein that we could remove the blue dye with ethanol, I
> think 90%. But the protein was freeze dried on the walls of a sample tube,
> not stuck to a PVDF membrane.
> Good luck with your project
> - Steve
> Steve Driska, Physiology Department, Temple University Medical School
> Philadelphia, PA 19140 USA (215) 707-3283
> driska at astro.ocis.temple.edu
Check the quality and age of your chemicals. When I did my PhD-thesis,
too many years ago, I had similar problems with our protein sequencer.
After changing to a new Coomassie and SDS batch of higher quality, the
If you do not want to buy better Coomassie, check Anal. Biochem. during
the last two, three years, or so; I have seen at least two papers about
prpearing higher quality Coomassie from usual commercial products.
More information about the Proteins