Jove
Visualize
Contact Us
JoVE
x logofacebook logolinkedin logoyoutube logo
ABOUT JoVE
OverviewLeadershipBlogJoVE Help Center
AUTHORS
Publishing ProcessEditorial BoardScope & PoliciesPeer ReviewFAQSubmit
LIBRARIANS
TestimonialsSubscriptionsAccessResourcesLibrary Advisory BoardFAQ
RESEARCH
JoVE JournalMethods CollectionsJoVE Encyclopedia of ExperimentsArchive
EDUCATION
JoVE CoreJoVE BusinessJoVE Science EducationJoVE Lab ManualFaculty Resource CenterFaculty Site
Terms & Conditions of Use
Privacy Policy
Policies

Related Concept Videos

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by
Same author

G6PD deficiency and the exportome of plasmodium falciparum FCB-2: a comparative analysis utilizing one-dimensional blue-native electrophoresis and timstof mass spectrometry.

Parasitology research·2026
Same author

Omics and Multiomics-Based Diagnostics for Invasive Candidiasis: Toward Precision Medicine.

Molecular & cellular proteomics : MCP·2025
Same author

Type-I interferons in Vulvovaginal Candidiasis: Mechanism of epithelial early defense and immune regulation against Candida albicans.

Mucosal immunology·2025
Same author

Integrative Phosphoproteomic and Proteomic Analysis of <i>Candida albicans</i> Exposed to Oxidative Stress.

Journal of proteome research·2025
Same author

<i>Candida albicans</i>: A Comprehensive View of the Proteome.

Journal of proteome research·2025
Same author

From High Protection to Lethal Effect: Diverse Outcomes of Immunization Against Invasive Candidiasis with Different <i>Candida albicans</i> Extracellular Vesicles.

International journal of molecular sciences·2025

Related Experiment Video

Updated: Aug 22, 2025

Quantitative Phosphoproteomics in Fatty Acid Stimulated Saccharomyces cerevisiae
15:41

Quantitative Phosphoproteomics in Fatty Acid Stimulated Saccharomyces cerevisiae

Published on: October 12, 2009

10.1K

SILAC-Based Quantitative Phosphoproteomics in Yeast.

María Luisa Hernáez1, Concha Gil2,3

  • 1Unidad de Proteómica, Facultad de Farmacia, Universidad Complutense de Madrid, Madrid, Spain.

Methods in Molecular Biology (Clifton, N.J.)
|November 12, 2022
PubMed
Summary

This study details Stable Isotope Labeling with Amino acids in Cell culture (SILAC) methods for yeast phosphoproteome analysis. Optimized procedures enable precise quantification of protein posttranslational modifications in signaling pathways.

Keywords:
In-solution digestionPhosphopeptides enrichmentPhosphoproteomicsQ-ExactiveQuantitative ProteomicsSILACTiO2

More Related Videos

Identification of protein complexes with quantitative proteomics in S. cerevisiae
11:12

Identification of protein complexes with quantitative proteomics in S. cerevisiae

Published on: March 4, 2009

13.3K
Quantitative Analysis of the Cellular Lipidome of Saccharomyces Cerevisiae Using Liquid Chromatography Coupled with Tandem Mass Spectrometry
08:56

Quantitative Analysis of the Cellular Lipidome of Saccharomyces Cerevisiae Using Liquid Chromatography Coupled with Tandem Mass Spectrometry

Published on: March 8, 2020

7.4K

Related Experiment Videos

Last Updated: Aug 22, 2025

Quantitative Phosphoproteomics in Fatty Acid Stimulated Saccharomyces cerevisiae
15:41

Quantitative Phosphoproteomics in Fatty Acid Stimulated Saccharomyces cerevisiae

Published on: October 12, 2009

10.1K
Identification of protein complexes with quantitative proteomics in S. cerevisiae
11:12

Identification of protein complexes with quantitative proteomics in S. cerevisiae

Published on: March 4, 2009

13.3K
Quantitative Analysis of the Cellular Lipidome of Saccharomyces Cerevisiae Using Liquid Chromatography Coupled with Tandem Mass Spectrometry
08:56

Quantitative Analysis of the Cellular Lipidome of Saccharomyces Cerevisiae Using Liquid Chromatography Coupled with Tandem Mass Spectrometry

Published on: March 8, 2020

7.4K

Area of Science:

  • Proteomics
  • Molecular Biology
  • Biochemistry

Background:

  • Quantitative proteomics is crucial for understanding cellular signaling.
  • Stable Isotope Labeling with Amino acids in Cell culture (SILAC) is a powerful technique for relative protein quantification.
  • Analyzing posttranslational modifications like phosphorylation requires robust enrichment and detection methods.

Purpose of the Study:

  • To describe detailed procedures for SILAC labeling in yeast.
  • To optimize and evaluate phosphopeptide enrichment strategies.
  • To establish a comprehensive workflow for global phosphoproteome analysis.

Main Methods:

  • Stable Isotope Labeling with Amino acids in Cell culture (SILAC) applied to yeast auxotrophs using labeled lysine and arginine.
  • Phosphopeptide enrichment using TiO2 chromatography.
  • High-resolution liquid chromatography-tandem mass spectrometry (LC-MS/MS) for identification and quantification.

Main Results:

  • Established a double SILAC method for yeast.
  • Optimized phosphopeptide enrichment protocols.
  • Enabled accurate quantification of phosphosites and protein abundance.

Conclusions:

  • The described SILAC workflow combined with phosphopeptide enrichment is effective for global phosphoproteome analysis.
  • This approach facilitates the study of signaling pathways through the measurement of protein posttranslational modifications.
  • The detailed procedures provide a valuable resource for researchers in yeast biology and signaling.