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磷酸浓缩结合无标签定量质谱法研究前列腺癌的 Phosphoproteome
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JoVE Journal
Ricerca sul cancro
Phosphopeptide Enrichment Coupled with Label-free Quantitative Mass Spectrometry to Investigate the Phosphoproteome in Prostate Cancer
Please note that all translations are automatically generated.
Click here for the English version.
磷酸浓缩结合无标签定量质谱法研究前列腺癌的 Phosphoproteome
DOI:
10.3791/57996-v
•
12:23 min
•
August 02, 2018
•
Larry C. Cheng*
1,2
,
Zhen Li*
3
,
Thomas G. Graeber
,
Nicholas A. Graham
,
Justin M. Drake
2,3,6,7
1
Graduate Program in Cellular and Molecular Pharmacology, School of Graduate Studies
,
Rutgers University, The State University of New Jersey
,
2
Graduate Program in Quantitative Biomedicine, School of Graduate Studies
,
Rutgers University, The State University of New Jersey
,
3
Department of Medicine, Division of Medical Oncology
,
Rutgers Robert Wood Johnson Medical School
,
4
Crump 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
,
5
Mork Family Department of Chemical Engineering and Materials Science
,
University of Southern California
,
6
Pharmacology
,
Rutgers Robert Wood Johnson Medical School
,
7
Cancer Metabolism and Growth Program
,
Rutgers Cancer Institute of New Jersey
Capitoli
00:04
Titolo
00:38
Harvesting Tissue
01:31
Lysate Digestion and Reverse Phase Extraction
04:12
Immunoprecipitation and Enrichment of pY Peptides and Prewashing of pY Beads
06:59
Titanium Dioxide Enrichment of pY Peptides and Desalting for MS Analyses
09:32
Results: Prostate Cancer Phosphopeptide Enrichment and Analysis by Mass Spectrometry
11:29
Conclusion
Summary
Traduzione automatica
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本协议描述了从前列腺癌细胞系或组织中提取和丰富磷酸肽持的过程,
通过
质谱法蛋白质组学分析 phosphoproteome。
Tags
Keywords: Phosphoproteome
Phosphopeptide Enrichment
Label-free Quantitative Mass Spectrometry
Prostate Cancer
Signaling Networks
Therapeutic Resistance
Lysis Buffer
Homogenization
Reduction
Alkylation
Sonication
Centrifugation
Trypsin Digestion
Filter
Acidification
C18 Column
Vacuum Manifold
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