TY - JOUR
T1 - A simple colorimetric method for the measurement of hydrogen peroxide produced by cells in culture
AU - Pick, Edgar
AU - Keisari, Yona
N1 - Funding Information:
1 Supported by a grant from the Deutsches Krebsforschungszentrum, Heidelberg, F.R.G. and by Grant No. 1505 from the U.S.-Israel Binational Science Foundation. 2 To whom reprint requests should be addressed.
PY - 1980
Y1 - 1980
N2 - A simple, rapid and inexpensive method for the measurement of hydrogen peroxide (H2O2) produced by cells in culture is described. The assay is based on the horseradish peroxidase of a (HRPO)-mediated oxidation of phenol red by H2O2 which results in the formation of a compound demonstrating increased absorbance at 610 nm. A linear relationship between absorbance at 610 nm and concentration of H2O2 was found in the 1–60 μM (1–60 nmoles/ml) range. Due to the non-toxic character of phenol red and HRPO, the assay permits measurement of H2O2 production and release by macrophages for time intervals of 5–60 min under regular tissue culture conditions. Using this assay, the ability of a number of agents to induce H2O2 release by guinea pig peritoneal macrophages was demonstrated. These agents were: phorbol myristate acetate (PMA), opsonized zymosan, concanavalin A (Con A), wheat germ agglutinin (WGA), N-formyl-methionyl-leucyl-phenylalanine (FMLP) and A23187.
AB - A simple, rapid and inexpensive method for the measurement of hydrogen peroxide (H2O2) produced by cells in culture is described. The assay is based on the horseradish peroxidase of a (HRPO)-mediated oxidation of phenol red by H2O2 which results in the formation of a compound demonstrating increased absorbance at 610 nm. A linear relationship between absorbance at 610 nm and concentration of H2O2 was found in the 1–60 μM (1–60 nmoles/ml) range. Due to the non-toxic character of phenol red and HRPO, the assay permits measurement of H2O2 production and release by macrophages for time intervals of 5–60 min under regular tissue culture conditions. Using this assay, the ability of a number of agents to induce H2O2 release by guinea pig peritoneal macrophages was demonstrated. These agents were: phorbol myristate acetate (PMA), opsonized zymosan, concanavalin A (Con A), wheat germ agglutinin (WGA), N-formyl-methionyl-leucyl-phenylalanine (FMLP) and A23187.
UR - http://www.scopus.com/inward/record.url?scp=0019140628&partnerID=8YFLogxK
U2 - 10.1016/0022-1759(80)90340-3
DO - 10.1016/0022-1759(80)90340-3
M3 - ???researchoutput.researchoutputtypes.contributiontojournal.article???
AN - SCOPUS:0019140628
SN - 0022-1759
VL - 38
SP - 161
EP - 170
JO - Journal of Immunological Methods
JF - Journal of Immunological Methods
IS - 1-2
ER -
