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A radioligand receptor binding assay for measuring of insulin secreted by MIN6 cells after stimulation with glucose, arginine, ornithine, dopamine, and serotonin
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SYSNO ASEP 0543107 Document Type J - Journal Article R&D Document Type Journal Article Subsidiary J Článek ve WOS Title A radioligand receptor binding assay for measuring of insulin secreted by MIN6 cells after stimulation with glucose, arginine, ornithine, dopamine, and serotonin Author(s) Asai, Seiya (UOCHB-X) ORCID
Žáková, Lenka (UOCHB-X) RID, ORCID
Selicharová, Irena (UOCHB-X) RID, ORCID
Marek, Aleš (UOCHB-X) RID, ORCID
Jiráček, Jiří (UOCHB-X) RID, ORCIDSource Title Analytical and Bioanalytical Chemistry. - : Springer - ISSN 1618-2642
Roč. 413, č. 17 (2021), s. 4531-4543Number of pages 13 s. Language eng - English Country DE - Germany Keywords binding assay ; insulin receptor ; insulin secretion ; radioligand ; secretagogue ; β Cells OECD category Biochemistry and molecular biology R&D Projects EF16_019/0000729 GA MŠMT - Ministry of Education, Youth and Sports (MEYS) Method of publishing Limited access Institutional support UOCHB-X - RVO:61388963 UT WOS 000655957900001 EID SCOPUS 85107001343 DOI 10.1007/s00216-021-03423-3 Annotation We adapted a radioligand receptor binding assay for measuring insulin levels in unknown samples. The assay enables rapid and accurate determination of insulin concentrations in experimental samples, such as from insulin-secreting cells. The principle of the method is based on the binding competition of insulin in a measured sample with a radiolabeled insulin for insulin receptor (IR) in IM-9 cells. Both key components, radiolabeled insulin and IM-9 cells, are commercially available. The IR binding assay was used to determine unknown amounts of insulin secreted by MIN6 β cell line after stimulation with glucose, arginine, ornithine, dopamine, and serotonin. The experimental data obtained by the IR binding assay were compared to the results determined by RIA kits and both methods showed a very good agreement of results. We observed the stimulation of glucose-induced insulin secretion from MIN6 cells by arginine, weaker stimulation by ornithine, but inhibitory effects of dopamine. Serotonin effects were either stimulatory or inhibitory, depending on the concentration of serotonin used. The results will require further investigation. The study also clearly revealed advantages of the IR binding assay that allows the measuring of a higher throughput of measured samples, with a broader range of concentrations than in the case of RIA kits. The IR binding assay can provide an alternative to standard RIA and ELISA assays for the determination of insulin levels in experimental samples and can be especially useful in scientific laboratories studying insulin production and secretion by β cells and searching for new modulators of insulin secretion. Workplace Institute of Organic Chemistry and Biochemistry Contact asep@uochb.cas.cz ; Kateřina Šperková, Tel.: 232 002 584 ; Viktorie Chládková, Tel.: 232 002 434 Year of Publishing 2022 Electronic address https://doi.org/10.1007/s00216-021-03423-3
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