In binding competition assays using a protein kinase C preparation from mouse brain (particulate fraction) 3H-labelled 12-O-tetradecanoylphorbol-13-acetate (TPA), for a series of new diterpene esters (DTE) the relative binding affinity [rba = Kia(TPA)/Kia(DTE)] in relation to TPA was determined. A wide range of values was noticed, some of the DTE binding more strongly than TPA (rba greater than 1), others binding less strongly than TPA (rba less than 1) In comparative terms, competition for specific binding sites appears to correlate better with irritant than with promoting activity of the DTE. Using mouse peritoneal neutrophils, binding of [3H]-TPA was determined by a modification of the "cold-acetone filter assay"; saturation of high-affinity sites (Kda = 0.2 nM) was obtained at concentrations less than or equal to 1 nM, but there was also evidence for specific binding at "low-affinity" sites (Kda = 26 nM). Induction of chemoluminescence in the presence of luminol in mouse peritoneal neutrophils with a set of DTE usually elecited two peaks; at concentrations greater than or equal to 10 nM DTE a short-lived, "spike-like" response lasting only from 0 to about 5 min (phase A) its followed by a "plateau" response from about 5-120 min (phase B). This latter phase of chemoluminescence stimulation with luminol correlated well with the irritant potential of the DTE used. The sequence of the two phases can be inverted partially by using first TPA at 2,5 nM followed by a quick concentration increase to 100 nM; this indicates two different concentration-dependent events. As regards the intensity of the chemoluminescent response, quantitative but not qualitative differences between DTE were observed, which show some correlation with strong and weak tumour-promoting activity. Inhibition studies suggest the involvement of the myeloperoxidase/H2O2/Cl- system in the luminogenic response; it is suggested that the release of hypochlorite or a closely related oxidant may be instrumental in tumour promotion.