The Use of Titanium Dioxide for Selective Enrichment of Phosphorylated Peptides.
(2016) In Methods in Molecular Biology 1355. p.135-146- Abstract
- Titanium dioxide (TiO2) has very high affinity for phosphopeptides and in recent years it has become one of the most popular methods for phosphopeptide enrichment from complex biological samples. Peptide loading onto TiO2 resin in a highly acidic environment in the presence of 2,5-dihydroxybenzoic acid (DHB), phthalic acid, lactic acid, or glycolic acid has been shown to improve selectivity significantly by reducing unspecific binding of non-phosphorylated peptides. The phosphopeptides bound to the TiO2 are subsequently eluted from the chromatographic material using an alkaline buffer. TiO2 chromatography is extremely tolerant towards most buffers used in biological experiments, highly robust and as such it has become the method of choice... (More)
- Titanium dioxide (TiO2) has very high affinity for phosphopeptides and in recent years it has become one of the most popular methods for phosphopeptide enrichment from complex biological samples. Peptide loading onto TiO2 resin in a highly acidic environment in the presence of 2,5-dihydroxybenzoic acid (DHB), phthalic acid, lactic acid, or glycolic acid has been shown to improve selectivity significantly by reducing unspecific binding of non-phosphorylated peptides. The phosphopeptides bound to the TiO2 are subsequently eluted from the chromatographic material using an alkaline buffer. TiO2 chromatography is extremely tolerant towards most buffers used in biological experiments, highly robust and as such it has become the method of choice in large-scale phosphoproteomics. Here we describe a batch mode protocol for phosphopeptide enrichment using TiO2 chromatographic material followed by desalting and concentration of the sample by reversed phase micro-columns prior to downstream MS and LC-MS/MS analysis. (Less)
Please use this url to cite or link to this publication:
https://lup.lub.lu.se/record/8235109
- author
- Thingholm, Tine LU and Larsen, Martin R
- organization
- publishing date
- 2016
- type
- Contribution to journal
- publication status
- published
- subject
- in
- Methods in Molecular Biology
- volume
- 1355
- pages
- 135 - 146
- publisher
- Springer
- external identifiers
-
- pmid:26584923
- scopus:84947976150
- pmid:26584923
- ISSN
- 1940-6029
- DOI
- 10.1007/978-1-4939-3049-4_9
- language
- English
- LU publication?
- yes
- id
- aef55d3f-8c23-4150-bf88-6cded2ae3da0 (old id 8235109)
- alternative location
- http://www.ncbi.nlm.nih.gov/pubmed/26584923?dopt=Abstract
- date added to LUP
- 2016-04-04 08:35:49
- date last changed
- 2022-04-23 17:43:09
@article{aef55d3f-8c23-4150-bf88-6cded2ae3da0,
abstract = {{Titanium dioxide (TiO2) has very high affinity for phosphopeptides and in recent years it has become one of the most popular methods for phosphopeptide enrichment from complex biological samples. Peptide loading onto TiO2 resin in a highly acidic environment in the presence of 2,5-dihydroxybenzoic acid (DHB), phthalic acid, lactic acid, or glycolic acid has been shown to improve selectivity significantly by reducing unspecific binding of non-phosphorylated peptides. The phosphopeptides bound to the TiO2 are subsequently eluted from the chromatographic material using an alkaline buffer. TiO2 chromatography is extremely tolerant towards most buffers used in biological experiments, highly robust and as such it has become the method of choice in large-scale phosphoproteomics. Here we describe a batch mode protocol for phosphopeptide enrichment using TiO2 chromatographic material followed by desalting and concentration of the sample by reversed phase micro-columns prior to downstream MS and LC-MS/MS analysis.}},
author = {{Thingholm, Tine and Larsen, Martin R}},
issn = {{1940-6029}},
language = {{eng}},
pages = {{135--146}},
publisher = {{Springer}},
series = {{Methods in Molecular Biology}},
title = {{The Use of Titanium Dioxide for Selective Enrichment of Phosphorylated Peptides.}},
url = {{http://dx.doi.org/10.1007/978-1-4939-3049-4_9}},
doi = {{10.1007/978-1-4939-3049-4_9}},
volume = {{1355}},
year = {{2016}},
}