Microspotting streptavidin and double-stranded DNA arrays on gold for high-throughput studies of protein-DNA interactions by surface plasmon resonance microscopy

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dc.contributor.author Zareie, HM
dc.contributor.author Shumaker-Parry, JS
dc.contributor.author Aebersold, R
dc.contributor.author Campbell, CT
dc.date.accessioned 2009-06-26T04:10:47Z
dc.date.issued 2004-01
dc.identifier.citation Analytical Chemistry, 2004, 76 (4), pp. 918 - 929
dc.identifier.issn 0003-2700
dc.identifier.other C1UNSUBMIT en_US
dc.identifier.uri http://hdl.handle.net/10453/515
dc.description.abstract We present two strategies for microspotting 10 × 12 arrays of double-stranded DNAs (dsDNAs) onto a gold-coated glass slide for high-throughput studies of protein-DNA interactions by surface plasmon resonance (SPR) microscopy. Both methods use streptavidin (SA) as a linker layer between a biotin-containing mixed self-assembled monolayer (SAM) and biotinylated dsDNAs to produce arrays with high packing density. The primary mixed SAM is produced from biotin- and oligo(ethylene glycol)-terminated thiols bonded as thiolates onto the gold surface. In the first method, a robotic microspotter is used to deliver nanoliter droplets of dsDNA solution onto a uniform layer of this SA (~2 × 1012 SA/cm2). SPR microscopy shows a density of (5-6) × 1011 dsDNA/cm2 (0.2-0.3 dsDNA/SA) in the array elements. The second method uses instead a microspotted array of this SA linker layer, onto which the microspots of dsDNA are added with spatial registry. SPR microscopy before addition of the dsDNA shows a SA coverage of 2 × 1012 SA/cm2 within the spots and a dsDNA density of 8.5 ± 3.5 × 1011 dsDNA/cm2 (0.3-0.7 dsDNA/SA, depending on the length of dsDNA) after dsDNA spotting. We demonstrate the ability to simultaneously monitor protein binding with the SPR microscope in many 200-m spots with 1-s time resolution and sensitivity to <1 pg of protein.
dc.publisher ACS Publications
dc.relation.isbasedon 10.1021/ac034964v
dc.title Microspotting streptavidin and double-stranded DNA arrays on gold for high-throughput studies of protein-DNA interactions by surface plasmon resonance microscopy
dc.type Journal Article
dc.parent Analytical Chemistry
dc.journal.volume 4
dc.journal.volume 76
dc.journal.number 4 en_US
dc.publocation Ohio, USA en_US
dc.identifier.startpage 918 en_US
dc.identifier.endpage 929 en_US
dc.cauo.name SCI.Physics and Advanced Materials en_US
dc.conference Verified OK en_US
dc.for 0301 Analytical Chemistry
dc.personcode 030414
dc.percentage 100 en_US
dc.classification.name Analytical Chemistry en_US
dc.classification.type FOR-08 en_US
pubs.embargo.period Not known
pubs.organisational-group /University of Technology Sydney
pubs.organisational-group /University of Technology Sydney/Faculty of Science
utslib.copyright.status Closed Access
utslib.copyright.date 2015-04-15 12:17:09.805752+10
utslib.copyright.date 2015-04-15 12:17:09.805752+10
pubs.consider-herdc false
pubs.consider-herdc false
utslib.collection.history General Collection (ID: 346) [2015-05-15T14:11:10+10:00]
utslib.collection.history School of Physics and Advanced Materials (ID: 343)
utslib.collection.history Closed (ID: 3)


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