{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,22]],"date-time":"2026-09-22T00:14:00Z","timestamp":1790036040398,"version":"4.0.1"},"reference-count":88,"publisher":"MDPI AG","issue":"24","license":[{"start":{"date-parts":[[2022,12,16]],"date-time":"2022-12-16T00:00:00Z","timestamp":1671148800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/100000002","name":"National Institutes of Health","doi-asserted-by":"publisher","award":["RF1NS113839"],"award-info":[{"award-number":["RF1NS113839"]}],"id":[{"id":"10.13039\/100000002","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000002","name":"National Institutes of Health","doi-asserted-by":"publisher","award":["SNF310030_ 207763"],"award-info":[{"award-number":["SNF310030_ 207763"]}],"id":[{"id":"10.13039\/100000002","id-type":"DOI","asserted-by":"publisher"}]},{"name":"Swiss National Science Foundation","award":["RF1NS113839"],"award-info":[{"award-number":["RF1NS113839"]}]},{"name":"Swiss National Science Foundation","award":["SNF310030_ 207763"],"award-info":[{"award-number":["SNF310030_ 207763"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>Calcitonin gene-related peptide (CGRP) is a key component of migraine pathophysiology, yielding effective migraine therapeutics. CGRP receptors contain a core accessory protein subunit: receptor activity-modifying protein 1 (RAMP1). Understanding of RAMP1 expression is incomplete, partly due to the challenges in identifying specific and validated antibody tools. We profiled antibodies for immunodetection of RAMP1 using Western blotting, immunocytochemistry and immunohistochemistry, including using RAMP1 knockout mouse tissue. Most antibodies could detect RAMP1 in Western blotting and immunocytochemistry using transfected cells. Two antibodies (844, ab256575) could detect a RAMP1-like band in Western blots of rodent brain but not RAMP1 knockout mice. However, cross-reactivity with other proteins was evident for all antibodies. This cross-reactivity prevented clear conclusions about RAMP1 anatomical localization, as each antibody detected a distinct pattern of immunoreactivity in rodent brain. We cannot confidently attribute immunoreactivity produced by RAMP1 antibodies (including 844) to the presence of RAMP1 protein in immunohistochemical applications in brain tissue. RAMP1 expression in brain and other tissues therefore needs to be revisited using RAMP1 antibodies that have been comprehensively validated using multiple strategies to establish multiple lines of convincing evidence. As RAMP1 is important for other GPCR\/ligand pairings, our results have broader significance beyond the CGRP field.<\/jats:p>","DOI":"10.3390\/ijms232416035","type":"journal-article","created":{"date-parts":[[2022,12,19]],"date-time":"2022-12-19T04:54:13Z","timestamp":1671425653000},"page":"16035","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":13,"title":["Characterization of Antibodies against Receptor Activity-Modifying Protein 1 (RAMP1): A Cautionary Tale"],"prefix":"10.3390","volume":"23","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-0763-5044","authenticated-orcid":false,"given":"Erica R.","family":"Hendrikse","sequence":"first","affiliation":[{"name":"School of Biological Sciences, The University of Auckland, Auckland 1010, New Zealand"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tayla A.","family":"Rees","sequence":"additional","affiliation":[{"name":"School of Biological Sciences, The University of Auckland, Auckland 1010, New Zealand"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7652-6004","authenticated-orcid":false,"given":"Zoe","family":"Tasma","sequence":"additional","affiliation":[{"name":"School of Biological Sciences, The University of Auckland, Auckland 1010, New Zealand"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5332-1236","authenticated-orcid":false,"given":"Michael L.","family":"Garelja","sequence":"additional","affiliation":[{"name":"Department of Pharmacology and Toxicology, The University of Otago, Dunedin 9016, New Zealand"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0341-065X","authenticated-orcid":false,"given":"Andrew","family":"Siow","sequence":"additional","affiliation":[{"name":"School of Biological Sciences, The University of Auckland, Auckland 1010, New Zealand"},{"name":"School of Chemical Sciences, The University of Auckland, Auckland 1010, New Zealand"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Paul W. R.","family":"Harris","sequence":"additional","affiliation":[{"name":"School of Biological Sciences, The University of Auckland, Auckland 1010, New Zealand"},{"name":"School of Chemical Sciences, The University of Auckland, Auckland 1010, New Zealand"},{"name":"Maurice Wilkins Centre for Molecular Biodiscovery, The University of Auckland, Auckland 1010, New Zealand"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2407-2944","authenticated-orcid":false,"given":"John B.","family":"Pawlak","sequence":"additional","affiliation":[{"name":"Department of Cell Biology and Physiology, University of North Carolina at Chapel Hill, Chapel Hill, NC 27599, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kathleen M.","family":"Caron","sequence":"additional","affiliation":[{"name":"Department of Cell Biology and Physiology, University of North Carolina at Chapel Hill, Chapel Hill, NC 27599, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Elizabeth S.","family":"Blakeney","sequence":"additional","affiliation":[{"name":"Department of Cell Biology and Physiology, University of North Carolina at Chapel Hill, Chapel Hill, NC 27599, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8156-5649","authenticated-orcid":false,"given":"Andrew F.","family":"Russo","sequence":"additional","affiliation":[{"name":"Department of Molecular Physiology and Biophysics, University of Iowa, Iowa City, IA 52242, USA"},{"name":"Center for the Prevention and Treatment of Visual Loss, Veterans Administration Health Center, Iowa City, IA 52246, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Levi P.","family":"Sowers","sequence":"additional","affiliation":[{"name":"Department of Molecular Physiology and Biophysics, University of Iowa, Iowa City, IA 52242, USA"},{"name":"Center for the Prevention and Treatment of Visual Loss, Veterans Administration Health Center, Iowa City, IA 52246, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5056-8548","authenticated-orcid":false,"given":"Thomas A.","family":"Lutz","sequence":"additional","affiliation":[{"name":"Institute of Veterinary Physiology, University of Zurich, Zurich 8057, Switzerland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6677-5488","authenticated-orcid":false,"given":"Christelle","family":"Le Foll","sequence":"additional","affiliation":[{"name":"Institute of Veterinary Physiology, University of Zurich, Zurich 8057, Switzerland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8151-4123","authenticated-orcid":false,"given":"Christopher S.","family":"Walker","sequence":"additional","affiliation":[{"name":"School of Biological Sciences, The University of Auckland, Auckland 1010, New Zealand"},{"name":"Maurice Wilkins Centre for Molecular Biodiscovery, The University of Auckland, Auckland 1010, New Zealand"}],"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, The University of Otago, Dunedin 9016, New Zealand"},{"name":"Maurice Wilkins Centre for Molecular Biodiscovery, The University of Auckland, Auckland 1010, New Zealand"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2022,12,16]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"96","DOI":"10.1186\/s10194-017-0807-1","article-title":"Blocking CGRP in migraine patients\u2014A review of pros and cons","volume":"18","author":"Deen","year":"2017","journal-title":"J. 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