A DISCONTINUOUS METHOD FOR CATALASE DETERMINATION AT NEAR PHYSIOLOGICAL CONCENTRATIONS OF H2O2 AND ITS APPLICATION TO THE STUDY OF H2O2 FLUXES WITHIN CELLS

Authors
OU PM WOLFF SP
Citation
Pm. Ou et Sp. Wolff, A DISCONTINUOUS METHOD FOR CATALASE DETERMINATION AT NEAR PHYSIOLOGICAL CONCENTRATIONS OF H2O2 AND ITS APPLICATION TO THE STUDY OF H2O2 FLUXES WITHIN CELLS, Journal of biochemical and biophysical methods, 31(1-2), 1996, pp. 59-67
Citations number
12
Categorie Soggetti
Biology,Biophysics,"Biochemical Research Methods
Journal title
Journal of biochemical and biophysical methods → ACNP
ISSN journal
0165022X
Volume
31
Issue
1-2
Year of publication
1996
Pages
59 - 67
Database
ISI
SICI code
0165-022X(1996)31:1-2<59:ADMFCD>2.0.ZU;2-F
Abstract
We describe a discontinuous method for the measurement of catalase usi ng the Ferrous Oxidation in Xylenol orange (FOX) assay. Samples contai ning catalase are incubated with H2O2 for varying time intervals prior to rapid mixing of aliquots of the incubation mixtures with FOX reage nt, which measures residual H2O2. Absorbance is then read at 560 nm af ter 30-min incubation at room temperature. Decay of H2O2 is proportion al to catalase activity in the original sample. An adaptation of the m ethod involves exposure of intact cells to H2O2-generating systems in the presence of aminotriazole. The extent of catalase inactivation all ows estimates of total H2O2 accumulation within the cell.