I used melatonin ELISA kits from IBL INTERNATIONAL GMBH and I didn't have parallelism nor a good standard curve. I need to measure melatonin levels in perch, pikeperch, rudd and roach.
We had in the past a radioimmunoassay for melatonin that worked well. It was used for sheep serum. The assay details and validation are published in:
Ronayne E., Jordan B., Quirke J.F., Roche J.F.
The effect of frequency of administration of melatonin on the time of onset of the breeding season in anoestrous ewes
(1989) Animal Reproduction Science, 18 (1-3) , pp. 13-24.
One of the most active groups working on melatonin in salmonid fish was the University of Stirling group of Neil Bromage, together with Mark Porter and others. The RIA protocol for their melatonin assay is found in the paper by Randall et al 1995, attached.
Thank you very much ! I dicussed my problem with Hervé Migaud from University of Stirling and they used this ELISA kit in the past with good results in salmonids but they actually prefer the RIA protocol.
I think that I have to switch to the RIA method too.
We've been using ELISA (after RIA) for many years; it works very well. Check this recent interesting paper from V Cassone for cheap easy to make ELISA at home : Li, Y.;Cassone, VM "A simple, specific high-throughput enzyme-linked immunosorbent assay (ELISA) for quantitative determination of melatonin in cell culture medium." International immunopharmacology 28, 1 (2015): 230-234.
HPLC also works well providing you have the appropriate equipment.
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