A microtiter plate-based assay for phosphoenolpyruvate carboxylase

A sensitive, quantitative assay for phosphenolpyruvate carboxylase which utilizes microtiter plates is described. The assay depends upon the production of a colored compound in the reaction between oxaloacetate, the product of the phosphoenolpyruvate reaction, and the dye Fast Violet B. The method i...

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Veröffentlicht in:	Analytical biochemistry 1990-08, Vol.189 (1), p.95-98
Hauptverfasser:	COCKBURN, W, WHITELAM, G. C, SLOCOMBE, S. P, MCKEE, R. A
Format:	Artikel
Sprache:	eng
Schlagworte:	Analytical, structural and metabolic biochemistry Biological and medical sciences Diazonium Compounds - chemistry Enzymes and enzyme inhibitors Fundamental and applied biological sciences. Psychology Hydrogen-Ion Concentration Lyases Microchemistry - methods Phosphoenolpyruvate Carboxylase - analysis Regression Analysis Reproducibility of Results Spectrophotometry - methods Staining and Labeling
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Beschreibung
Zusammenfassung:	A sensitive, quantitative assay for phosphenolpyruvate carboxylase which utilizes microtiter plates is described. The assay depends upon the production of a colored compound in the reaction between oxaloacetate, the product of the phosphoenolpyruvate reaction, and the dye Fast Violet B. The method is particularly appropriate for monitoring chromatographic eluates and its utility for this purpose is demonstrated by the detection of phosphoenolpyruvate carboxylase in fractions of crude maize extract separated by size-exclusion chromatography.
ISSN:	0003-2697 1096-0309
DOI:	10.1016/0003-2697(90)90051-A