An enzyme-linked immunosorbent assay (ELISA) for bovine LH capable of monitoring fluctuations in baseline concentrations

Using a Fab'-horseradish peroxidase conjugate, an ELISA for bovine LH was developed and validated. It had good specificity (less than or equal to 0.2% cross-reactivity with FSH or TSH), was not susceptible to interference from plasma components and exhibited very low non-specific binding. It was carried out on microtitre plates and had incubation times totalling 4 or 18 h, not including the initial antibody coating step. The longer incubations gave a lower detection limit (10 pg, 300 amol) and were used when fluctuations in baseline concentrations were being monitored. The values determined were independent of the plasma volume employed, standard added to plasma samples was accurately determined and the results obtained from the analysis of plasma samples correlated closely with those obtained by radioimmunoassay. The assay was applied to the analysis of plasma samples taken from heifers with normal ovarian activity or treated in a number of ways, and in every case the results obtained were similar to those determined by radioimmunoassay and reported in the literature.