Measurement of endogenous and TNFα-mediated H2O2 production in supernatants of SK-N-SH neuroblastoma cells with an enhanced chemiluminescence assay
✍ Scribed by Carmine, T. C. ;Bruchelt, G. ;Hahn, T. ;Niethammer, D.
- Book ID
- 101717137
- Publisher
- John Wiley and Sons
- Year
- 1994
- Weight
- 434 KB
- Volume
- 9
- Category
- Article
- ISSN
- 0884-3996
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✦ Synopsis
Abstract
A sensitive peroxidase‐dependent luminol‐enhanced chemiluminescence (ECL) assay for determination of hydrogen peroxide (H~2~O~2~) generation by tumour cells was established. This test system allows determination of H~2~O~2~ 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 10^4^SK‐N‐SH neuroblastoma cells released 60° 5 pmol H~2~O~2~ in the supernatant and this level was significantly (p<0.025) increased by about 70% in the presence of 5pmol (100 ng/mL) recombinant tumour necrosis factor α (TNFα). In contrast, H~2~O~2~ production was slightly reduced by TNFα at a very low concentration of 0.5 fmol (0.01 ng/mL).