METHODS article

Front. Mol. Biosci.

Sec. Protein Biochemistry for Basic and Applied Sciences

Volume 12 - 2025 | doi: 10.3389/fmolb.2025.1505250

This article is part of the Research TopicCellular Contributors and Consequences of Protein Misfolding and AggregationView all 9 articles

Thioflavin T In-gel Stain to Study Cardiac Amyloid Ex Vivo

Provisionally accepted
Joseph OldamJoseph Oldam1Irina TchernyshyovIrina Tchernyshyov2Jennifer Van EykJennifer Van Eyk1,2Juan TroncosoJuan Troncoso1Charles GlabeCharles Glabe3Giulio AgnettiGiulio Agnetti1,4*
  • 1Johns Hopkins University, Baltimore, Maryland, United States
  • 2Cedars Sinai Medical Center, Los Angeles, California, United States
  • 3University of California, Irvine, Irvine, California, United States
  • 4University of Bologna, Bologna, Italy

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There are limited options to quantify and characterize amyloid species from biological samples in a simple fashion. Thioflavin T (ThT) has now been used for decades to stain amyloid fibrils but to our knowledge we were the first to use it ingel. Thioflavin T in-gel stain is convenient as it is fast, inexpensive, available to most labs, compatible with other fluorescent stains and downstream analyses such as mass spectrometry (MS).Organ and systemic proteinopathies represent one of the main areas of public health concern in Westernized society in this century 1 . The toxic nature of prefibrillar and fibrillar amyloid is well-established in neurocognitive disease 2 , however their role and prevalence are emerging in cardiovascular disease as well 3 . Most amyloid species are quantitated by either fluorescent, radiolabeled, or immunological probes as well as by mass spectrometry (MS) [4][5][6][7] . From the technological standpoint, MS is arguably one of the techniques with the highest impact on biomedical sciences in this century 8 . Cutting-edge MS-based approaches such as isotope incorporation are currently being used in the study of protein misfolding but their application is still confined to a few laboratories worldwide 9 . It is our opinion that a simple way to detect and quickly isolate amyloid Deleted: Glabe 2 30 Deleted: 3

Keywords: Amyloid, Mass Spectrometry, proteinopathies, Desmin, Heart Failure

Received: 02 Oct 2024; Accepted: 14 Apr 2025.

Copyright: © 2025 Oldam, Tchernyshyov, Van Eyk, Troncoso, Glabe and Agnetti. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

* Correspondence: Giulio Agnetti, University of Bologna, Bologna, Italy

Disclaimer: All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article or claim that may be made by its manufacturer is not guaranteed or endorsed by the publisher.

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