Optimization of enzyme-linked immunosorbent assay kit protocol to detect trimethylamine N-oxide levels in humans

Abstract: The serum level of trimethylamine N-oxide (TMAO), a gut microbiota metabolite associated with diabetes, cancer, inflammatory and neurological diseases, can be determined by the micro-enzyme-linked immunosorbent assay (ELISA) method. However, we had problems obtaining accurate standard curves with the original kit protocol from Bioassay Technology Laboratory. We aimed to acquire proper standard curves by modifying the kit protocol in this study. First, we evaluated the human TMAO ELISA kit protocols and other human ELISA kits. We maintained the incubation times longer and increased the wash cycle. Moreover, we incubated the standards containing biotinylated antibody in the wells alone. Then we washed the wells and added streptavidin-HRP for the second incubation step. The data of original and modified ELISA kit protocol were analyzed with Student's t-test. We measured higher absorbance with lower standard solution concentration in experiments that followed the original kit protocol..... https://www.excli.de/index.php/excli/article/view/5617

Object Details

• Location: Deutsche Nationalbibliothek Frankfurt am Main
• Extent: Online-Ressource
• Language: Englisch

References and Relationships

• Bibliographic citation: Optimization of enzyme-linked immunosorbent assay kit protocol to detect trimethylamine N-oxide levels in humans ; volume:22 ; year:2023 ; EXCLI journal ; 22 (2023)

Contributors, Places and Time

• Creator: Aksoyalp, Zinnet Sevval ; Erdoğan, Betül Rabia ; Aksun, Saliha

Further information

• DOI: 10.17179/excli2022-5617
• URN: urn:nbn:de:101:1-2023033103142128477725
• Rights: Open Access; Der Zugriff auf das Objekt ist unbeschränkt möglich.
• Last update: 14.08.2025, 10:54 AM CEST