Phosphopeptide Enrichment Coupled with Label-free Quantitative Mass Spectrometry to Investigate the Phosphoproteome in Prostate Cancer
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Cited by 15
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12:23 min
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August 2nd, 2018
DOI :10.3791/57996-v
August 2nd, 2018
•14K views
Larry C. Cheng*1,2 , Zhen Li*3 , Thomas G. Graeber4 , Nicholas A. Graham5 , Justin M. Drake1,2,3,6,7
1Graduate Program in Cellular and Molecular Pharmacology, School of Graduate Studies, Rutgers University, The State University of New Jersey, 2Graduate Program in Quantitative Biomedicine, School of Graduate Studies, Rutgers University, The State University of New Jersey, 3Department of Medicine, Division of Medical Oncology, Rutgers Robert Wood Johnson Medical School, 4Crump Institute for Molecular Imaging, Department of Molecular and Medical Pharmacology, Jonsson Comprehensive Cancer Center, UCLA Metabolomics Center, and California NanoSystems Institute, David Geffen School of Medicine, University of California, Los Angeles, 5Mork Family Department of Chemical Engineering and Materials Science, University of Southern California, 6Pharmacology, Rutgers Robert Wood Johnson Medical School, 7Cancer Metabolism and Growth Program, Rutgers Cancer Institute of New Jersey
* These authors contributed equally
This protocol describes a procedure to extract and enrich phosphopeptides from prostate cancer cell lines or tissues for an analysis of the phosphoproteome via mass spectrometry-based proteomics.
Chapters in this video
0:04
Title
0:38
Harvesting Tissue
1:31
Lysate Digestion and Reverse Phase Extraction
4:12
Immunoprecipitation and Enrichment of pY Peptides and Prewashing of pY Beads
6:59
Titanium Dioxide Enrichment of pY Peptides and Desalting for MS Analyses
9:32
Results: Prostate Cancer Phosphopeptide Enrichment and Analysis by Mass Spectrometry
11:29
Conclusion
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