A simple, specific high-throughput enzyme-linked immunosorbent assay (ELISA) for quantitative determination of melatonin in cell culture medium

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28 Scopus citations

Abstract

A simple, specific, high-throughput enzyme-linked immunosorbent assay (ELISA) for quantitative determination of melatonin was developed for directly measuring melatonin in cell culture medium with 10% FBS. This assay adopts a commercial monoclonal melatonin antibody and melatonin-HRP conjugate, so it can be applied in multiple labs rapidly with low cost compared with commercial RIA and ELISA kits. In addition, the procedure is much simpler with only four steps: 1) sample/conjugate incubation, 2) plate washing, 3) TMB color reaction and 4) reading of results. The standards of the assay cover a wide working range from 100 pg/mL to 10 ng/mL. The sensitivity was 68 pg/mL in cell culture medium with 10% FBS and 26 pg/mL in PBS with as little as 25 μL sample volume. The recovery of melatonin from cell culture medium was 101.0%. The principal cross-reacting compound was 5-methoxytryptophol (0.1%). The variation coefficients of the assay, within and between runs, ranged between 6.68% and 15.76% in cell culture medium. The mean linearity of a series diluted cell culture medium sample was 105% (CV = 5%), ranging between 98% and 111%, y = 5.5263x + 0.0646, R2 = 0.99. The assay enables small research and teaching labs to reliably measure this important neurohormone.

Original languageEnglish
Pages (from-to)230-234
Number of pages5
JournalInternational Immunopharmacology
Volume28
Issue number1
DOIs
StatePublished - Jun 22 2015

Bibliographical note

Publisher Copyright:
© 2015 Elsevier B.V. All rights reserved.

Keywords

  • Chicken pineal
  • ELISA
  • Melatonin
  • Melatonin-HRP conjugate

ASJC Scopus subject areas

  • Immunology and Allergy
  • Immunology
  • Pharmacology

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