How about staining with amido black instead?
Ian mc
In article <5f4s21$vse at cronkite.ocis.temple.edu>,
driska at astro.ocis.temple.edu (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
