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dc.contributor.authorKilcoyle, Michelle
dc.contributor.authorGerlach, Jared Q.
dc.contributor.authorFarrell, Mark P.
dc.contributor.authorJoshi, Lokesh
dc.date.accessioned2013-09-04T12:16:55Z
dc.date.available2013-09-04T12:16:55Z
dc.date.issued2011
dc.identifier.citationKilcoyne, M,Gerlach, JQ,Farrell, MP,Bhavanandan, VP,Joshi, L (2011) 'Periodic acid-Schiff's reagent assay for carbohydrates in a microtiter plate format'. Analytical Biochemistry, 416 :18-26.en_US
dc.identifier.urihttp://hdl.handle.net/10379/3641
dc.description.abstractMicrotiter 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 25 Mu-l of sample, utilises standardised Schiff's reagent, and has decreased assay time (140 min 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 10-100 nmol range approximately, which varied for each carbohydrate. The assay response for fetuin and porcine gastric mucin (PGM) was linear up to 150 mu-g (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.en_US
dc.formatapplication/pdfen_US
dc.language.isoenen_US
dc.relation.ispartofAnalytical Biochemistryen
dc.subjectPAS assayen_US
dc.subjectSchiff's reagenten_US
dc.subjectPeriodateen_US
dc.subjectMucinsen_US
dc.subjectGlycoproteinsen_US
dc.subjectCOLORIMETRIC DETERMINATIONen_US
dc.subjectGASTROINTESTINAL MUCINSen_US
dc.subjectO-GLYCOSYLATIONen_US
dc.subjectPROTEINSen_US
dc.subjectGLYCOPROTEINSen_US
dc.subjectFETUINen_US
dc.subjectSUGARSen_US
dc.subjectRATen_US
dc.titlePeriodic acid-Schiff's reagent assay for carbohydrates in a microtiter plate formaten_US
dc.typeArticleen_US
dc.date.updated2013-07-24T12:19:20Z
dc.identifier.doiDOI 10.1016/j.ab.2011.05.006
dc.local.publishedsourcehttp://dx.doi.org/10.1016/j.ab.2011.05.006en_US
dc.description.peer-reviewedpeer-reviewed
dc.contributor.funder|~|
dc.internal.rssid1126888
dc.local.contactLokesh Joshi, School Of Natural Sciences, Nui Galway. 5768 Email: lokesh.joshi@nuigalway.ie
dc.local.copyrightcheckedYes
dc.local.versionACCEPTED
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