| Simultaneous capillary electrophoresis competitive immunoassay for insulin, glucagon, and islet amyloid polypeptide secretion from mouse islets of Langerhans. | |
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MedLine Citation:
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PMID: 21620410 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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A capillary electrophoresis competitive immunoassay was developed for the simultaneous quantitation of insulin, glucagon, and islet amyloid polypeptide (IAPP) secretion from islets of Langerhans. Separation buffers and conditions were optimized for the resolution of fluorescein isothiocyanate (FITC)-labeled glucagon and IAPP immunoassay reagents, which were excited with the 488 nm line of an Ar(+) laser and detected at 520 nm with a photomultiplier tube (PMT). Cy5-labeled insulin immunoassay reagents were excited by a 635 nm laser diode module and detected at 700 nm with a separate PMT. Optimum resolution was achieved with a 20mM carbonate separation buffer at pH 9.0 using a 20 cm effective separation length with an electric field of 500 V/cm. Limits of detection for insulin, glucagon, and IAPP were 2, 3, and 3 nM, respectively. This method was used to monitor the simultaneous secretion of these peptides from as few as 14 islets after incubation in 4, 11, and 20 mM glucose for 6h. For insulin and IAPP, a statistically significant increase in secretion levels was observed, while glucagon levels were significantly reduced in the 4 and 11 mM glucose conditions. To further demonstrate the utility of the assay, the Ca(2+)-dependent secretion of these peptides was demonstrated which agreed with published reports. The ability to examine the secretion of multiple peptides may allow for the determination of regulation of secretory processes within islets of Langerhans. |
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Authors:
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Christelle Guillo; Tuan M Truong; Michael G Roper |
Publication Detail:
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Type: Journal Article; Research Support, N.I.H., Extramural Date: 2011-05-13 |
Journal Detail:
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Title: Journal of chromatography. A Volume: 1218 ISSN: 1873-3778 ISO Abbreviation: J Chromatogr A Publication Date: 2011 Jul |
Date Detail:
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Created Date: 2011-06-06 Completed Date: 2011-09-01 Revised Date: 2011-09-26 |
Medline Journal Info:
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Nlm Unique ID: 9318488 Medline TA: J Chromatogr A Country: Netherlands |
Other Details:
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Languages: eng Pagination: 4059-64 Citation Subset: IM |
Copyright Information:
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Copyright © 2011 Elsevier B.V. All rights reserved. |
Affiliation:
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Department of Chemistry and Biochemistry, Florida State University, Tallahassee, FL 32306, USA. |
Export Citation:
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| MeSH Terms | |
Descriptor/Qualifier:
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Animals Binding, Competitive Calcium Calibration Electrophoresis, Capillary / methods* Glucagon / analysis*, secretion Immunoassay / methods* Insulin / analysis*, secretion Islet Amyloid Polypeptide / analysis*, secretion Islets of Langerhans Male Mice Rats Reproducibility of Results Sensitivity and Specificity Swine |
| Grant Support | |
ID/Acronym/Agency:
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R01 DK080714/DK/NIDDK NIH HHS; R01 DK080714-05/DK/NIDDK NIH HHS |
| Chemical | |
Reg. No./Substance:
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0/Islet Amyloid Polypeptide; 11061-68-0/Insulin; 7440-70-2/Calcium; 9007-92-5/Glucagon |
| Comments/Corrections | |
Erratum In:
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J Chromatogr A. 2011 Aug 19;1218(33):5731 Note: Truong, Tuan M [added] |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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