In solution digestion - (Mar/12/2013 )

Hi,

I am doing my in solution digestion with trypsine. And this is my pilot study with BSA.
According to optimal pH for trypsine activity I have tried to adjust the solution pH to 8-8.5. Usually I use ammonium bicarbonate 50mM for adjusting. but this time I use Ammonia. Now I want to know if it will work or the ammonia will have some effect on the procedure and also for peptides?

Review a general chemistry textbook. Ammonia and Ammonia bicarbonate are very similiar, sometime regarded as analogous in chemical reactions. Ammonia with form ammonia bicarbonate in the presence of water and carbon dioxide. The ammonia you add, will ultimately form ammonia bicarbonate (depending on your reaction, but more than likely). It wont have any affect on your protein. As far as I know, ammonia salts will be formed following digestion of individual amino acids.

Edit: reference. Unfortunately, this is the only reference I can find. I know I remember reading about ammonia forming ammonia salts following amino acid cleavage (probably in voet and voet).

http://cancerres.aacrjournals.org/content/38/8/2199.full.pdf

thanks a lot