Cytokines, insulin-like growth factor-1, PGE2 and NO determination

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Procedure

Specific ELISAs for TNF-α, IL-6, IL-10 (Endogen Inc.), IL-4, insulin-like growth
factor-1 (IGF-1, Boster Biological Technology) and high sensitivity EIA for PGE2
(Assay Designs) were used to analyse OHSC culture media, following the
manufacturer’s instructions. The ranges of determination were: 31-2500 pg/ml for TNF-
α, 31-2000 pg/ml for IL-6; 7.8 pg/ml-500 pg/ml for IL-4; 16-500 pg/ml for IL-10; 31-
2000 pg/ml for IGF-1; 7.8-1000 pg/ml for PGE2.
The production of nitric oxide (NO) was determined by measuring the content of nitrite,
one of the end products of NO oxidation in the media, as previously described
(Minghetti et al., 1996). A standard nitrite curve (0.25-50 μM) was generated using a 10
mM solution of NaNO2. All chemical for the NO assay were from Sigma.

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