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dc.contributor.authorSchrader, S
dc.contributor.authorTiralongo, E
dc.contributor.authorParis, G
dc.contributor.authorYoshino, T
dc.contributor.authorSchauer, R
dc.date.accessioned2017-05-03T14:35:26Z
dc.date.available2017-05-03T14:35:26Z
dc.date.issued2003
dc.identifier.issn0003-2697
dc.identifier.doi10.1016/j.ab.2003.07.016
dc.identifier.urihttp://hdl.handle.net/10072/24068
dc.description.abstractTrans-sialidase (E.C. 3.2.1.18) catalyzes the transfer of preferably a2,3-linked sialic acid to another glycan or glycoconjugate, forming a new a2,3 linkage to galactose or N-acetylgalactosamine. Here, we describe a nonradioactive 96-well plate fluorescence test for monitoring trans-sialidase activity with high sensitivity, specificity, and reproducibility using sialyllactose and 4-methylumbelliferyl-߭Image -galactoside as donor and acceptor substrates, respectively. The assay conditions were optimized using the trans-sialidase from Trypanosoma congolense and its general applicability was confirmed with recombinant trans-sialidase from Trypanosoma cruzi. Using this procedure, a large number of samples can be tested quickly and reliably, for instance in monitoring trans-sialidase during enzyme purification and the production of monoclonal antibodies, for enzyme characterization, and for identifying potential substrates and inhibitors. The trans-sialidase assay reported here was capable of detecting trans-sialidase activity in the low-mU range and may be a valuable tool in the search for further trans-sialidases in various biological systems.
dc.description.peerreviewedYes
dc.description.publicationstatusYes
dc.languageEnglish
dc.language.isoeng
dc.publisherAcademic Press
dc.publisher.placeUSA
dc.relation.ispartofpagefrom139
dc.relation.ispartofpageto147
dc.relation.ispartofjournalAnalytical Biochemistry
dc.relation.ispartofvolume322
dc.subject.fieldofresearchAnalytical chemistry
dc.subject.fieldofresearchOther chemical sciences
dc.subject.fieldofresearchBiochemistry and cell biology
dc.subject.fieldofresearchcode3401
dc.subject.fieldofresearchcode3499
dc.subject.fieldofresearchcode3101
dc.titleA nonradioactive 96-well plate assay for screening of trans-sialidase activity
dc.typeJournal article
dc.type.descriptionC1 - Articles
dc.type.codeC - Journal Articles
gro.date.issued2015-02-06T01:36:46Z
gro.hasfulltextNo Full Text
gro.griffith.authorTiralongo, Evelin


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