Simple procedure for removal of AMP from NADP preparation.
A simple and reliable procedure for removal of AMP from NADP preparation is described. In this procedure, a mixture of AMP and NADP solution is first incubated with 5'-nucleotidase to hydrolyze AMP to adenosine and inorganic phosphate (Pi). The reaction mixture is then applied to a Dowex 1 (formate) column. Adenosine and 5'-nucleotidase are removed by washing the column with 20 mM HCOOH. NADP is finally eluted with 3.5 M HCOOH followed by precipitation and washing with acetone. The yield of salt-free NADP is about 80%. Although Pi is coeluted with NADP in the acid form (H3PO4), it is removed during the precipitation and repeated washing with acetone. A slight modification of this procedure for simultaneous removal of AMP, ADP, and ATP from NADP preparation has also been discussed.
D000249:Adenosine Monophosphate / Q000302:isolation & purification*; D000818:Animals; D002645:Chickens; D004340:Drug Contamination; D006597:Fructose-Bisphosphatase / Q000037:antagonists & inhibitors; D009132:Muscles / Q000201:enzymology; D009249:NADP / Q000032:analysis*
D000249:Adenosine Monophosphate; D009249:NADP; D006597:Fructose-Bisphosphatase