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 a

SUBMITTER: Aksoyalp ZS 

PROVIDER: S-EPMC10415589 | biostudies-literature | 2023

REPOSITORIES: biostudies-literature