| Periodic acid-Schiff's reagent assay for carbohydrates in a microtiter plate format. | |
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MedLine Citation:
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PMID: 21620792 Owner: NLM Status: Publisher |
Abstract/OtherAbstract:
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Microtiter plate colorimetric assays are widely used for analysis of carbohydrates and glycoconjugates. However, mucins are often not easily detected, as they have low neutral sugar content. We have adapted and optimised the periodic acid-Schiff's reagent (PAS) staining for microtiter plate assay by examining five factors: concentration and volume of periodic acid, oxidation time, volume of Schiff's reagent, and color development time. This assay requires just 25μl of sample, utilises standardised Schiff's reagent, and has decreased assay time (140min to completion). Seventeen monosaccharides (acidic, neutral, basic, phosphorylated, and deoxy) and four disaccharides were assessed. PAS-positive carbohydrates (amino, N-acetylamino, deoxy, and certain neutral monosaccharides, and sialic acids) responded linearly within a 10-100nmol range approximately, which varied for each carbohydrate. The assay response for fetuin and porcine gastric mucin (PGM) was linear up to 150μg (highest concentration tested), with no response from nonglycosylated protein. A lower response for asialofetuin was observed, but desialylated PGM preparations were similar or higher in response than their sialylated counterparts. The simplicity and low sample consumption of this method make it an excellent choice for screening or quantitation of chromatographic fractions containing carbohydrates and glycoconjugates, especially in the case of mucins. |
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Authors:
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Michelle Kilcoyne; Jared Q Gerlach; Mark P Farrell; Veer P Bhavanandan; Lokesh Joshi |
Publication Detail:
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Type: JOURNAL ARTICLE Date: 2011-5-10 |
Journal Detail:
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Title: Analytical biochemistry Volume: - ISSN: 1096-0309 ISO Abbreviation: - Publication Date: 2011 May |
Date Detail:
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Created Date: 2011-5-30 Completed Date: - Revised Date: - |
Medline Journal Info:
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Nlm Unique ID: 0370535 Medline TA: Anal Biochem Country: - |
Other Details:
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Languages: ENG Pagination: - Citation Subset: - |
Copyright Information:
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Copyright © 2011 Elsevier Inc. All rights reserved. |
Affiliation:
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Glycoscience Group, National Centre for Biomedical Engineering Science, National University of Ireland, Galway, Ireland. |
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From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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