Amylin antibodies frequently display cross-reactivity with CGRP: characterization of eight amylin antibodies

Author:

Rees Tayla A.12ORCID,Hay Debbie L.123ORCID,Walker Christopher S.12ORCID

Affiliation:

1. School of Biological Sciences, University of Auckland, Auckland, New Zealand

2. Maurice Wilkins Centre for Molecular Biodiscovery, University of Auckland, Auckland, New Zealand

3. Department of Pharmacology and Toxicology, University of Otago, Dunedin, New Zealand

Abstract

Amylin is a 37-amino acid endocrine hormone secreted from the pancreas in response to nutrient intake, acting centrally to promote meal-ending satiation. With many studies linking amylin action to the nervous system, determining the distribution or expression of amylin in the nervous system is critical. However, amylin shares sequence identity and structural homology to the related neuropeptide calcitonin gene-related peptide (CGRP). This creates challenges in identifying selective amylin antibodies that do not cross-react with CGRP, especially in neural tissues, where CGRP is densely packed into secretory vesicles. Here, we characterized eight amylin antibodies to determine their ability to detect amylin and cross-react with rat or human αCGRP, using immunoblots and preabsorption controls in rat pancreas. We observed that amylin antibodies frequently cross-reacted with αCGRP and are therefore not suitable for use in tissues that highly express CGRP. Earlier work using these antibodies should be revisited in light of our findings.

Funder

Manatu Hauora | Health Research Council of New Zealand

Royal Society of New Zealand

Royal Society of New Zealand | Marsden Fund

University of Auckland

Publisher

American Physiological Society

Subject

Physiology (medical),Physiology

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