Photometric microtiter assay of inorganic phosphate in the presence of acid-labile organic phosphates

Analytical Biochemistry
P DrueckesD Palm

Abstract

A simple and rapid colorimetric microassay for inorganic phosphate in the mild acid pH range has enabled us to perform extensive enzyme kinetic studies even in the presence of high concentrations of acid-labile substrates. The assay is performed in a 96-well microtiter plate using 30-microliters samples containing between 0.5 and 100 nmol Pi. Compared to existing microassays, color formation is linear over a much wider range of phosphate concentrations and reaction conditions. Using a computer-assisted microtiter plate reader, the data can be directly transferred to kinetic analysis software. The above assay was used to determine simultaneously up to 12 rates of the Escherichia coli maltodextrin phosphorylase-catalyzed reaction in the direction of glycogen synthesis under conditions which include screening of substrates and inhibitors at variable substrate concentrations and pH on a single 96-well microtiter plate.

Citations

Jan 15, 2005·Pharmaceutical Research·Elena V BatrakovaAlexander V Kabanov
Sep 29, 2006·Antimicrobial Agents and Chemotherapy·Adrian S RayGerry R Rhodes
May 13, 1999·Proceedings of the National Academy of Sciences of the United States of America·M E EtzlerJ B Murphy
Dec 31, 2009·Journal of Environmental and Public Health·Takaharu Mizutani
Mar 29, 2008·Expert Opinion on Drug Metabolism & Toxicology·James J CaliJohn Shultz
Sep 25, 2001·Xenobiotica; the Fate of Foreign Compounds in Biological Systems·S AbelD K Walker
Feb 15, 2011·Phytochemistry·Abbas M Lafta, Karen Klotz Fugate
May 16, 2009·European Journal of Pharmaceutical Sciences : Official Journal of the European Federation for Pharmaceutical Sciences·Saijie ZhuYuanying Pei
Jul 1, 2005·Protein Science : a Publication of the Protein Society·Nikos G OikonomakosElisabeth Defossa
Mar 23, 2004·Analytical Biochemistry·Jeffrey D Gawronski, David R Benson
Mar 2, 2010·Analytical Biochemistry·M R M De GroeveW Soetaert
Jul 10, 2003·Biochemical and Biophysical Research Communications·Daniel BallezaFroylan Gómez-Lagunas
Jul 18, 2006·Drug Metabolism and Drug Interactions·Thomas Broeng EjsingKristian Linnet
Feb 9, 2017·Journal of Molecular Microbiology and Biotechnology·Hanna S RuppersbergRalf Rabus
Oct 21, 2004·Analytical Biochemistry·Ana D SimonovićMarc D Anderson
Sep 7, 2004·Journal of Clinical Psychopharmacology·Sebastian HärtterChristoph Hiemke
Oct 26, 2007·Archives of Pharmacal Research·Hye Young JiHye Suk Lee
Feb 28, 2006·The Journal of Biological Chemistry·Kei TakahashiKazumitsu Ueda
Jan 28, 2010·Dalton Transactions : an International Journal of Inorganic Chemistry·Cindy A PuckettJacqueline K Barton
Oct 27, 2007·Drug Metabolism and Disposition : the Biological Fate of Chemicals·Bo FengSonia M de Morais
May 25, 2004·The Journal of Biological Chemistry·Guillaume LenoirPierre Falson
Sep 3, 2002·Advanced Drug Delivery Reviews·Alexander V KabanovValery Yu Alakhov
Sep 30, 2005·Journal of Medicinal Chemistry·Thomas KlabundeElisabeth Defossa
Mar 29, 2003·Toxicology and Applied Pharmacology·Nilufer M TampalGabriele Ludewig

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