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Biochemistry
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通过 FASP 消化和数据独立分析,EPS-尿液的蛋白质配置文件
JoVE Journal
Biochemistry
Author Produced
This content is Free Access.
JoVE Journal
Biochemistry
Proteomic Profile of EPS-Urine through FASP Digestion and Data-Independent Analysis
Please note that all translations are automatically generated.
Click here for the English version.
通过 FASP 消化和数据独立分析,EPS-尿液的蛋白质配置文件
DOI:
10.3791/62512-v
•
14:48 min
•
May 08, 2021
•
Licia E. Prestagiacomo
,
Caterina Gabriele
,
Paola Morelli
,
Maria Antonietta Rota
,
Stefano Alba
,
Giovanni Cuda
,
Rocco Damiano
,
Marco Gaspari
1
Research Centre for Advanced Biochemistry and Molecular Biology, Department of Experimental and Clinical Medicine
,
Magna Graecia University of Catanzaro
,
2
Department of Health Science
,
Magna Graecia University of Catanzaro
,
3
Romolo Hospital
,
4
Department of Experimental and Clinical Medicine
,
Magna Graecia University of Catanzaro
Chapters
00:00
Introduction
00:55
Protocol
01:25
Protocol: FASP Digestion
05:29
Protocol: SCX Purification
08:08
Protocol: Protein Quantifcation by External Standard
09:23
Protocol: NanoLC-MS/MS and DIA Analysis
11:31
Protocol: High-pH C18 Fractionation for Spectral Library
13:09
Protocol: Data Analysis
13:35
Expected Results
13:53
Conclusion
Summary
Automatic Translation
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Automatic Translation
在这里,我们提出了一个优化的过滤消化协议,并提供了以下详细信息:蛋白质消化、肽纯化和数据独立获取分析。该策略应用于对表达的前列腺分泌物-尿液样本的分析,并允许高蛋白体覆盖率和低缺失值无标签尿素分析。
Tags
Proteomic Profile
EPS-urine
FASP Digestion
Data-independent Analysis
Mass Spectrometry
Biomarker Discovery
Prostate Cancer
Centrifugation
Sample Preparation
Protein Alkylation
Urea Buffer
Iodoacetamide
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