(my sample is protein bar formulated with dates apricots cheese and whey protein Isolate. I am interested to study the protein and peptides in my product through electrophrosis. So, please let me know how can i prepare my sample for protein analysis and how can i make it free from sucrose and fructose)
Thank you
Dear Sidra,
I suggest that dialysis is a suitable method to get rid of the monosaccharides.
Best wishes, Peter
Hi, Sidra
Ammonium sulfate precipitation followed by dialysis is suitable to purify the proteins.
The CHCl3/MeOH method of Wessel & Flügge (doi:10.1016/0003-2697(84)90782-6) precipitates proteins and collects them in a film that redisolves well in SDS sample buffer. The method is quick, simple, cheap and quantitative, and it doesn't add salts that interfere with electrophoresis. However, it will not work for smaller peptides. But for those electrophoresis is unsuitable as well. For those I'd hydrolyse (6 M HCl, 120 °C, o/n) and then do amino acid analysis by ion exchange chromatography.
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