dc.contributor.author | Tape, CJ | |
dc.contributor.author | Worboys, JD | |
dc.contributor.author | Sinclair, J | |
dc.contributor.author | Gourlay, R | |
dc.contributor.author | Vogt, J | |
dc.contributor.author | McMahon, KM | |
dc.contributor.author | Trost, M | |
dc.contributor.author | Lauffenburger, DA | |
dc.contributor.author | Lamont, DJ | |
dc.contributor.author | Jørgensen, C | |
dc.date.accessioned | 2016-11-23T11:26:59Z | |
dc.date.issued | 2014-09-18 | |
dc.identifier.citation | Analytical chemistry, 2014, 86 (20), pp. 10296 - 10302 | |
dc.identifier.issn | 0003-2700 | |
dc.identifier.uri | https://repository.icr.ac.uk/handle/internal/235 | |
dc.identifier.eissn | 1520-6882 | |
dc.identifier.doi | 10.1021/ac5025842 | |
dc.description.abstract | Reproducible, comprehensive phosphopeptide enrichment is essential for studying phosphorylation-regulated processes. Here, we describe the application of hyper-porous magnetic TiO2 and Ti-IMAC microspheres for uniform automated phosphopeptide enrichment. Combining magnetic microspheres with a magnetic particle-handling robot enables rapid (45 min), reproducible (r2 ≥ 0.80) and high-fidelity (>90% purity) phosphopeptide purification in a 96-well format. Automated phosphopeptide enrichment demonstrates reproducible synthetic phosphopeptide recovery across 2 orders of magnitude, "well-to-well" quantitative reproducibility indistinguishable to internal SILAC standards, and robust "plate-to-plate" reproducibility across 5 days of independent enrichments. As a result, automated phosphopeptide enrichment enables statistical analysis of label-free phosphoproteomic samples in a high-throughput manner. This technique uses commercially available, off-the-shelf components and can be easily adopted by any laboratory interested in phosphoproteomic analysis. We provide a free downloadable automated phosphopeptide enrichment program to facilitate uniform interlaboratory collaboration and exchange of phosphoproteomic data sets. | |
dc.format | Print-Electronic | |
dc.format.extent | 10296 - 10302 | |
dc.language | eng | |
dc.language.iso | eng | |
dc.publisher | AMER CHEMICAL SOC | |
dc.subject | Titanium | |
dc.subject | Imidazoles | |
dc.subject | Phosphopeptides | |
dc.subject | Multivariate Analysis | |
dc.subject | Reproducibility of Results | |
dc.subject | Proteomics | |
dc.subject | Magnetics | |
dc.subject | Automation | |
dc.title | Reproducible automated phosphopeptide enrichment using magnetic TiO2 and Ti-IMAC. | |
dc.type | Journal Article | |
rioxxterms.versionofrecord | 10.1021/ac5025842 | |
rioxxterms.licenseref.startdate | 2014-10-02 | |
rioxxterms.type | Journal Article/Review | |
dc.relation.isPartOf | Analytical chemistry | |
pubs.issue | 20 | |
pubs.notes | No embargo | |
pubs.organisational-group | /ICR | |
pubs.organisational-group | /ICR/Primary Group | |
pubs.organisational-group | /ICR/Primary Group/ICR Divisions | |
pubs.organisational-group | /ICR/Primary Group/ICR Divisions/Closed research teams | |
pubs.organisational-group | /ICR/Primary Group/ICR Divisions/Closed research teams/Cell Communication | |
pubs.organisational-group | /ICR/Primary Group/ICR Divisions/Closed research teams/Oncogene | |
pubs.organisational-group | /ICR | |
pubs.organisational-group | /ICR/Primary Group | |
pubs.organisational-group | /ICR/Primary Group/ICR Divisions | |
pubs.organisational-group | /ICR/Primary Group/ICR Divisions/Closed research teams | |
pubs.organisational-group | /ICR/Primary Group/ICR Divisions/Closed research teams/Cell Communication | |
pubs.organisational-group | /ICR/Primary Group/ICR Divisions/Closed research teams/Oncogene | |
pubs.publication-status | Published | |
pubs.volume | 86 | |
pubs.embargo.terms | No embargo | |
icr.researchteam | Cell Communication | |
icr.researchteam | Oncogene | |
dc.contributor.icrauthor | Worboys, Jonathan David | |