We have investigated enzyme-linked immunosorbent assay (ELISA) as a possible alternative to radioimmunoassay (RIA) for the detection and measurement of the neuropeptide substance P (SP) and its metabolite substance P1-7. The sensitivities were higher with ELISA than with RIA utilizing the same antisera. The intra-assay and inter-assay variation in ELISA was 1% and 13% respectively. The higher sensitivities are in part due to the standard curves having less steep slopes, and in part to lower IC50s in the ELISA. Since there was a good correlation between peptide levels in biological samples as determined by ELISA and RIA respectively, ELISA might be considered an attractive alternative to RIA.