SILAC-Based Temporal Phosphoproteomics

Research output: Chapter in Book/Report/Conference proceeding › Book chapter › Education

Standard

SILAC-Based Temporal Phosphoproteomics. / Francavilla, Chiara; Hekmat, Omid; Blagoev, Blagoy; Olsen, Jesper V.

Stable Isotope Labeling by Amino Acids in Cell Culture (SILAC): Methods and Protocols. ed. / Bettina Warscheid. Vol. 1188 2014. p. 125-48 (Methods in molecular biology (Clifton, N.J.)).

Research output: Chapter in Book/Report/Conference proceeding › Book chapter › Education

Harvard

Francavilla, C, Hekmat, O, Blagoev, B & Olsen, JV 2014, SILAC-Based Temporal Phosphoproteomics. in B Warscheid (ed.), Stable Isotope Labeling by Amino Acids in Cell Culture (SILAC): Methods and Protocols. vol. 1188, Methods in molecular biology (Clifton, N.J.), pp. 125-48. https://doi.org/10.1007/978-1-4939-1142-4_10

APA

Francavilla, C., Hekmat, O., Blagoev, B., & Olsen, J. V. (2014). SILAC-Based Temporal Phosphoproteomics. In B. Warscheid (Ed.), Stable Isotope Labeling by Amino Acids in Cell Culture (SILAC): Methods and Protocols (Vol. 1188, pp. 125-48). Methods in molecular biology (Clifton, N.J.) https://doi.org/10.1007/978-1-4939-1142-4_10

Vancouver

Francavilla C, Hekmat O, Blagoev B, Olsen JV. SILAC-Based Temporal Phosphoproteomics. In Warscheid B, editor, Stable Isotope Labeling by Amino Acids in Cell Culture (SILAC): Methods and Protocols. Vol. 1188. 2014. p. 125-48. (Methods in molecular biology (Clifton, N.J.)). https://doi.org/10.1007/978-1-4939-1142-4_10

Author

Francavilla, Chiara ; Hekmat, Omid ; Blagoev, Blagoy ; Olsen, Jesper V. / SILAC-Based Temporal Phosphoproteomics. Stable Isotope Labeling by Amino Acids in Cell Culture (SILAC): Methods and Protocols. editor / Bettina Warscheid. Vol. 1188 2014. pp. 125-48 (Methods in molecular biology (Clifton, N.J.)).

Bibtex

@inbook{15223a9427d448e28e93873e18209383,
title = "SILAC-Based Temporal Phosphoproteomics",
abstract = "In recent years, thanks to advances in Mass Spectrometry (MS)-based quantitative proteomics, studies on signaling pathways have moved from a detailed description of individual components to system-wide analysis of entire signaling cascades, also providing spatio-temporal views of intracellular pathways. Quantitative proteomics that combines stable isotope labeling by amino acid in cell culture (SILAC) with enrichment strategies for post-translational modification-bearing peptides and high-performance tandem mass spectrometry represents a powerful and unbiased approach to monitor dynamic signaling events. Here we provide an optimized SILAC-based proteomic workflow to analyze temporal changes in phosphoproteomes, which involve a generic three step enrichment protocol for phosphopeptides. SILAC-labeled peptides from digested whole cell lysates are as a first step enriched for phosphorylated tyrosines by immunoaffinity and then further enriched for phosphorylated serine/threonine peptides by strong cation exchange in combination with titanium dioxide-beads chromatography. Analysis of enriched peptides on Orbitrap-based MS results in comprehensive and accurate reconstruction of temporal changes of signaling networks.",
author = "Chiara Francavilla and Omid Hekmat and Blagoy Blagoev and Olsen, {Jesper V}",
year = "2014",
doi = "10.1007/978-1-4939-1142-4_10",
language = "English",
isbn = "978-1-4939-1141-7",
volume = "1188",
series = "Methods in molecular biology (Clifton, N.J.)",
publisher = "Humana Press",
pages = "125--48",
editor = "Warscheid, {Bettina }",
booktitle = "Stable Isotope Labeling by Amino Acids in Cell Culture (SILAC)",

}

RIS

TY - CHAP

T1 - SILAC-Based Temporal Phosphoproteomics

AU - Francavilla, Chiara

AU - Hekmat, Omid

AU - Blagoev, Blagoy

AU - Olsen, Jesper V

PY - 2014

Y1 - 2014

N2 - In recent years, thanks to advances in Mass Spectrometry (MS)-based quantitative proteomics, studies on signaling pathways have moved from a detailed description of individual components to system-wide analysis of entire signaling cascades, also providing spatio-temporal views of intracellular pathways. Quantitative proteomics that combines stable isotope labeling by amino acid in cell culture (SILAC) with enrichment strategies for post-translational modification-bearing peptides and high-performance tandem mass spectrometry represents a powerful and unbiased approach to monitor dynamic signaling events. Here we provide an optimized SILAC-based proteomic workflow to analyze temporal changes in phosphoproteomes, which involve a generic three step enrichment protocol for phosphopeptides. SILAC-labeled peptides from digested whole cell lysates are as a first step enriched for phosphorylated tyrosines by immunoaffinity and then further enriched for phosphorylated serine/threonine peptides by strong cation exchange in combination with titanium dioxide-beads chromatography. Analysis of enriched peptides on Orbitrap-based MS results in comprehensive and accurate reconstruction of temporal changes of signaling networks.

AB - In recent years, thanks to advances in Mass Spectrometry (MS)-based quantitative proteomics, studies on signaling pathways have moved from a detailed description of individual components to system-wide analysis of entire signaling cascades, also providing spatio-temporal views of intracellular pathways. Quantitative proteomics that combines stable isotope labeling by amino acid in cell culture (SILAC) with enrichment strategies for post-translational modification-bearing peptides and high-performance tandem mass spectrometry represents a powerful and unbiased approach to monitor dynamic signaling events. Here we provide an optimized SILAC-based proteomic workflow to analyze temporal changes in phosphoproteomes, which involve a generic three step enrichment protocol for phosphopeptides. SILAC-labeled peptides from digested whole cell lysates are as a first step enriched for phosphorylated tyrosines by immunoaffinity and then further enriched for phosphorylated serine/threonine peptides by strong cation exchange in combination with titanium dioxide-beads chromatography. Analysis of enriched peptides on Orbitrap-based MS results in comprehensive and accurate reconstruction of temporal changes of signaling networks.

U2 - 10.1007/978-1-4939-1142-4_10

DO - 10.1007/978-1-4939-1142-4_10

M3 - Book chapter

C2 - 25059609

SN - 978-1-4939-1141-7

VL - 1188

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

SP - 125

EP - 148

BT - Stable Isotope Labeling by Amino Acids in Cell Culture (SILAC)

A2 - Warscheid, Bettina

ER -

ID: 119771428