Determination of horseradish peroxidase concentration using the chemiluminescence of Cypridina luciferin analogue, 2-methyl-6-(p-methoxyphenyl)-3,7-dihydroimidazo[1,2-a]pyrazin-3-one.
The chemiluminescence of the Cypridina luciferin analogue, 2-methyl-6-(p-methoxyphenyl)-3,7-dihydroimidazo[1,2-a]pyrazin++ +-3-one (MCLA) was observed at 462 nm in the presence of horseradish peroxidase (HRP) and the total spectrum of light emitted was found to depend linearly on HRP concentration. Methods for the determination of HRP concentration using the chemiluminescence was investigated. HRP could be detected in the range from 100 pmol/L to 100 nmol/L under the optimum condition, H2O2 (10 mmol/L) and MCLA (10 mumol/L) at pH 5.8.
Mitani
M
M
Tsukuba Research Laboratory, NOF Corporation, Ibaraki, Japan.
Yokoyama
Y
Y
Ichikawa
S
S
Sawada
H
H
Matsumoto
T
T
Fujimori
K
K
Kosugi
M
M
D000818:Animals; D049454:Firefly Luciferin / Q000031:analogs & derivatives*; D006735:Horseradish Peroxidase / Q000032:analysis*; D006861:Hydrogen Peroxide; D006863:Hydrogen-Ion Concentration; D007093:Imidazoles; D007118:Immunoassay / Q000379:methods; D008163:Luminescent Measurements; D011719:Pyrazines; D013050:Spectrometry, Fluorescence; D013053:Spectrophotometry
D007093:Imidazoles; D011719:Pyrazines; C063412:2-methyl-6-(4-methoxyphenyl)-3,7-dihydroimidazo(1,2-alpha)pyrazin-3-one; D049454:Firefly Luciferin; D006861:Hydrogen Peroxide; D006735:Horseradish Peroxidase