{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,8]],"date-time":"2026-09-08T17:14:20Z","timestamp":1788887660959,"version":"build-2803163510"},"reference-count":33,"publisher":"Wiley","issue":"10","license":[{"start":{"date-parts":[[2025,8,7]],"date-time":"2025-08-07T00:00:00Z","timestamp":1754524800000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by-nc\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100009193","name":"Marsden Fund","doi-asserted-by":"publisher","id":[{"id":"10.13039\/501100009193","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100009997","name":"International Headache Society","doi-asserted-by":"publisher","id":[{"id":"10.13039\/100009997","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["headachejournal.onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Headache"],"published-print":{"date-parts":[[2025,11]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:sec>\n                    <jats:title>Objectives\/Background<\/jats:title>\n                    <jats:p>This study was undertaken to test the ability of a widely used enzyme\u2010linked immunosorbent assay (ELISA) kit to detect calcitonin gene\u2010related peptide (CGRP) isoforms to better understand currently published clinical data. There is significant interest in measuring CGRP as a biomarker in headache and migraine research, with ELISAs being the preferred detection method. ELISAs use antibodies that have been raised against an antigen to allow selective quantification of an analyte in a sample. Understanding the specificity of these antibodies is crucial to interpreting results. One commercially available kit (Cusabio CSB\u2010E08210h, Kit A) is purported to specifically detect \u03b2\u2010CGRP (one of the two CGRP isoforms) over \u03b1\u2010CGRP and has been used to investigate the potential for CGRP to be used as a biomarker for migraine. We investigated the ability of this kit to detect multiple isoforms of CGRP to better interpret published clinical results. We used a second ELISA kit (Bertin Bioreagent, A05481, Kit B) that is nonselective for the different CGRP isoforms as a control to ensure the CGRP we used could be detected in ELISAs.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Methods<\/jats:title>\n                    <jats:p>We performed ELISAs according to the manufacturer's instructions, testing concentrations within the advertised range of each kit. At least three independent experiments were conducted for each kit.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Results<\/jats:title>\n                    <jats:p>Kit B was able to detect both human and mouse \u03b1\u2010CGRP and \u03b2\u2010CGRP with a high degree of reproducibility. In contrast, Kit A did not detect bioactive forms of human \u03b1\u2010CGRP or \u03b2\u2010CGRP, nor mouse \u03b1\u2010CGRP or \u03b2\u2010CGRP.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Conclusion<\/jats:title>\n                    <jats:p>Kit A may not be reliable for future studies, as it does not appear to detect mature bioactive CGRP. Importantly, conclusions from previous studies that used this kit may need to be reevaluated, as it is not clear what analyte the kit has detected. Our findings also highlight the importance of understanding research tools to ensure accurate interpretation of results.<\/jats:p>\n                  <\/jats:sec>","DOI":"10.1111\/head.15011","type":"journal-article","created":{"date-parts":[[2025,8,7]],"date-time":"2025-08-07T10:52:23Z","timestamp":1754563943000},"page":"1744-1753","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":19,"title":["Calcitonin gene\u2010related peptide and headache: Comparison of two commonly used assay kits highlights the perils of measuring neuropeptides with enzyme\u2010linked immunosorbent assays"],"prefix":"10.1111","volume":"65","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-5332-1236","authenticated-orcid":false,"given":"Michael L.","family":"Garelja","sequence":"first","affiliation":[{"name":"Department of Pharmacology and Toxicology University of Otago  Dunedin New Zealand"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tayla A.","family":"Rees","sequence":"additional","affiliation":[{"name":"Headache Group, Wolfson Sensory, Pain, and Regeneration Centre, Institute of Psychiatry, Psychology, and Neuroscience King's College London  London UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9558-5122","authenticated-orcid":false,"given":"Debbie L.","family":"Hay","sequence":"additional","affiliation":[{"name":"Department of Pharmacology and Toxicology University of Otago  Dunedin New 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