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Adapting Homogeneous Enzyme-Linked Competitive Binding Assays to Microtiter Plates

dc.contributor.authorKim B. ,en_US
dc.contributor.authorBuckwalter J. M. ,en_US
dc.contributor.authorMeyerhoff M. E. ,en_US
dc.date.accessioned2006-04-10T18:13:35Z
dc.date.available2006-04-10T18:13:35Z
dc.date.issued1994-04en_US
dc.identifier.citationKim B., , Buckwalter J. M., , Meyerhoff M. E., (1994/04)."Adapting Homogeneous Enzyme-Linked Competitive Binding Assays to Microtiter Plates." Analytical Biochemistry 218(1): 14-19. <http://hdl.handle.net/2027.42/31647>en_US
dc.identifier.urihttp://www.sciencedirect.com/science/article/B6W9V-45P0J40-58/2/8704e5ae7d49321c78a8d97ce6cfba12en_US
dc.identifier.urihttps://hdl.handle.net/2027.42/31647
dc.identifier.urihttp://www.ncbi.nlm.nih.gov/sites/entrez?cmd=retrieve&db=pubmed&list_uids=8053547&dopt=citationen_US
dc.description.abstractRecently devised homogeneous enzyme-linked binding assays useful for the rapid detection of carbohydrate structure/content of intact glycoproteins (via use of lectins as binders) and for quantitating given vitamins (e.g., biotin; using soluble binding proteins) are adapted successfully to a microtiter plate reader format. The problem of nonspecific adsorption of the binders and enzyme-saccharide/vitamin conjugates is solved via the addition of Tween 20 to the assay buffer. More convenient and reliable photometric detection of the preferred labeling enzyme, glucose-6-phosphate dehydrogenase (G6PDH), is accomplished by monitoring the rate of generation of reduced thio-NAD (from thio-NAD) at 405 nm instead of NADH (from NAD) at 340 nm. By employing these modifications it is shown that homogeneous enzyme-linked binding assays can be readily adapted to microtiter plates without loss in analytical assay performance. Results further suggest that other homogeneous assays based on G6PDH, including commercial EMIT assays used routinely in clinical chemistry laboratories for detecting drugs of abuse, could, in principle, be run on microtiter plates to significantly enhance sample throughput.en_US
dc.format.extent542894 bytes
dc.format.extent3118 bytes
dc.format.mimetypeapplication/pdf
dc.format.mimetypetext/plain
dc.language.isoen_US
dc.publisherElsevieren_US
dc.titleAdapting Homogeneous Enzyme-Linked Competitive Binding Assays to Microtiter Platesen_US
dc.typeArticleen_US
dc.rights.robotsIndexNoFollowen_US
dc.subject.hlbsecondlevelPublic Healthen_US
dc.subject.hlbsecondlevelChemistryen_US
dc.subject.hlbsecondlevelChemical Engineeringen_US
dc.subject.hlbsecondlevelBiological Chemistryen_US
dc.subject.hlbtoplevelEngineeringen_US
dc.subject.hlbtoplevelScienceen_US
dc.subject.hlbtoplevelHealth Sciencesen_US
dc.description.peerreviewedPeer Revieweden_US
dc.contributor.affiliationumDepartment of Chemistry, University of Michigan, Ann Arbor, MI, USA.en_US
dc.contributor.affiliationumDepartment of Chemistry, University of Michigan, Ann Arbor, MI, USA.en_US
dc.contributor.affiliationumDepartment of Chemistry, University of Michigan, Ann Arbor, MI, USA.en_US
dc.identifier.pmid8053547en_US
dc.description.bitstreamurlhttp://deepblue.lib.umich.edu/bitstream/2027.42/31647/1/0000581.pdfen_US
dc.identifier.doihttp://dx.doi.org/10.1006/abio.1994.1135en_US
dc.identifier.sourceAnalytical Biochemistryen_US
dc.owningcollnameInterdisciplinary and Peer-Reviewed


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