Measurement of endogenous and TNFα‐mediated H2O2 production in supernatants of SK‐N‐SH neuroblastoma cells with an enhanced chemiluminescence assay

Abstract
A sensitive peroxidase‐dependent luminol‐enhanced chemiluminescence (ECL) assay for determination of hydrogen peroxide (H2O2) generation by tumour cells was established. This test system allows determination of H2O2 in concentrations as low as 25 pmol (50nmol/L) and yields results which are comparable to those obtained using a less sensitive photometric method and a previously described scopoletin flurorescence assay. After 3h incubation time 104SK‐N‐SH neuroblastoma cells released 60° 5 pmol H2O2 in the supernatant and this level was significantly (p2O2 production was slightly reduced by TNFα at a very low concentration of 0.5 fmol (0.01 ng/mL).