Offline High pH Reversed-Phase Peptide Fractionation for Deep Phosphoproteome Coverage

Research output: Chapter in Book/Report/Conference proceeding › Book chapter › Research › peer-review

Standard

Offline High pH Reversed-Phase Peptide Fractionation for Deep Phosphoproteome Coverage. / Batth, Tanveer S; Olsen, Jesper V.

Phospho-Proteomics: Methods and Protocols. ed. / Louise von Stechow. Vol. 1355 Springer, 2016. p. 179-92 (Methods in molecular biology (Clifton, N.J.)).

Research output: Chapter in Book/Report/Conference proceeding › Book chapter › Research › peer-review

Harvard

Batth, TS & Olsen, JV 2016, Offline High pH Reversed-Phase Peptide Fractionation for Deep Phosphoproteome Coverage. in LV Stechow (ed.), Phospho-Proteomics: Methods and Protocols. vol. 1355, Springer, Methods in molecular biology (Clifton, N.J.), pp. 179-92. https://doi.org/10.1007/978-1-4939-3049-4_12

APA

Batth, T. S., & Olsen, J. V. (2016). Offline High pH Reversed-Phase Peptide Fractionation for Deep Phosphoproteome Coverage. In L. V. Stechow (Ed.), Phospho-Proteomics: Methods and Protocols (Vol. 1355, pp. 179-92). Springer. Methods in molecular biology (Clifton, N.J.) https://doi.org/10.1007/978-1-4939-3049-4_12

Vancouver

Batth TS, Olsen JV. Offline High pH Reversed-Phase Peptide Fractionation for Deep Phosphoproteome Coverage. In Stechow LV, editor, Phospho-Proteomics: Methods and Protocols. Vol. 1355. Springer. 2016. p. 179-92. (Methods in molecular biology (Clifton, N.J.)). https://doi.org/10.1007/978-1-4939-3049-4_12

Author

Batth, Tanveer S ; Olsen, Jesper V. / Offline High pH Reversed-Phase Peptide Fractionation for Deep Phosphoproteome Coverage. Phospho-Proteomics: Methods and Protocols. editor / Louise von Stechow. Vol. 1355 Springer, 2016. pp. 179-92 (Methods in molecular biology (Clifton, N.J.)).

Bibtex

@inbook{898f36cb5e1749ab921cb8734292dad5,
title = "Offline High pH Reversed-Phase Peptide Fractionation for Deep Phosphoproteome Coverage",
abstract = "Protein phosphorylation, a process in which kinases modify serines, threonines, and tyrosines with phosphoryl groups is of major importance in eukaryotic biology. Protein phosphorylation events are key initiators of signaling responses which determine cellular outcomes after environmental and metabolic stimuli, and are thus highly regulated. Therefore, studying the mechanism of regulation by phosphorylation, and pinpointing the exact site of phosphorylation on proteins is of high importance. This protocol describes in detail a phosphoproteomics workflow for ultra-deep coverage by fractionating peptide mixtures based on high pH (basic) reversed-phase chromatography prior to phosphopeptide enrichment and mass spectrometric analysis. Peptides are separated on a C18 reversed-phase column under basic conditions and fractions collected in timed intervals followed by concatenation of the fractions. Each Fraction is subsequently enriched for phosphopeptides using TiO2 followed by LC/MS analysis.",
author = "Batth, {Tanveer S} and Olsen, {Jesper V}",
note = "AR2016",
year = "2016",
doi = "10.1007/978-1-4939-3049-4_12",
language = "English",
isbn = "978-1-4939-3048-7",
volume = "1355",
series = "Methods in molecular biology (Clifton, N.J.)",
publisher = "Springer",
pages = "179--92",
editor = "Stechow, {Louise von}",
booktitle = "Phospho-Proteomics",
address = "Switzerland",

}

RIS

TY - CHAP

T1 - Offline High pH Reversed-Phase Peptide Fractionation for Deep Phosphoproteome Coverage

AU - Batth, Tanveer S

AU - Olsen, Jesper V

N1 - AR2016

PY - 2016

Y1 - 2016

N2 - Protein phosphorylation, a process in which kinases modify serines, threonines, and tyrosines with phosphoryl groups is of major importance in eukaryotic biology. Protein phosphorylation events are key initiators of signaling responses which determine cellular outcomes after environmental and metabolic stimuli, and are thus highly regulated. Therefore, studying the mechanism of regulation by phosphorylation, and pinpointing the exact site of phosphorylation on proteins is of high importance. This protocol describes in detail a phosphoproteomics workflow for ultra-deep coverage by fractionating peptide mixtures based on high pH (basic) reversed-phase chromatography prior to phosphopeptide enrichment and mass spectrometric analysis. Peptides are separated on a C18 reversed-phase column under basic conditions and fractions collected in timed intervals followed by concatenation of the fractions. Each Fraction is subsequently enriched for phosphopeptides using TiO2 followed by LC/MS analysis.

AB - Protein phosphorylation, a process in which kinases modify serines, threonines, and tyrosines with phosphoryl groups is of major importance in eukaryotic biology. Protein phosphorylation events are key initiators of signaling responses which determine cellular outcomes after environmental and metabolic stimuli, and are thus highly regulated. Therefore, studying the mechanism of regulation by phosphorylation, and pinpointing the exact site of phosphorylation on proteins is of high importance. This protocol describes in detail a phosphoproteomics workflow for ultra-deep coverage by fractionating peptide mixtures based on high pH (basic) reversed-phase chromatography prior to phosphopeptide enrichment and mass spectrometric analysis. Peptides are separated on a C18 reversed-phase column under basic conditions and fractions collected in timed intervals followed by concatenation of the fractions. Each Fraction is subsequently enriched for phosphopeptides using TiO2 followed by LC/MS analysis.

U2 - 10.1007/978-1-4939-3049-4_12

DO - 10.1007/978-1-4939-3049-4_12

M3 - Book chapter

C2 - 26584926

SN - 978-1-4939-3048-7

VL - 1355

T3 - Methods in molecular biology (Clifton, N.J.)

SP - 179

EP - 192

BT - Phospho-Proteomics

A2 - Stechow, Louise von

PB - Springer

ER -

ID: 173364797