DICP OpenIR
Subject Area物理化学
In Situ Sample Processing Approach (iSPA) for Comprehensive Quantitative Phosphoproteome Analysis
Huang, Junfeng1,2; Qin, Hongqiang1; Dong, Jing1; Song, Chunxia1; Bian, Yangyang1,2; Dong, Mingming1,2; Cheng, Kai1,2; Wang, Fangjun1; Sun, Deguang3; Wang, Liming3; Ye, Mingliang1; Zou, Hanfa1; Ye ML(叶明亮); Zou HF(邹汉法)
KeywordIn Situ Sample processIng Approach Phosphoprotein Enrichment On-beads Digestion Dimethyl Labeling Solid Phase Labeling Phosphoproteome Quantification Human Liver Hepatocellular Carcinoma
Source PublicationJOURNAL OF PROTEOME RESEARCH
2014-09-01
DOI10.1021/pr500454g
Volume13Issue:9Pages:3896-3904
Indexed BySCI
SubtypeArticle
WOS HeadingsScience & Technology ; Life Sciences & Biomedicine
WOS SubjectBiochemical Research Methods
WOS Research AreaBiochemistry & Molecular Biology
WOS KeywordION AFFINITY-CHROMATOGRAPHY ; PHOSPHORYLATED PROTEINS ; MASS-SPECTROMETRY ; SELECTIVE ENRICHMENT ; PHOSPHOPEPTIDES ; QUANTIFICATION ; LIVER ; IDENTIFICATION ; ADSORBENT ; PEPTIDES
AbstractCurrent sample preparation protocols for quantitative phosphoproteome analysis are tedious and time-consuming. Here, a facile in situ sample processing approach (iSPA) is developed by using macroporous Ti(IV)-IMAC microspheres as the preparation "beds", where all sample preparation procedures including the enrichment of phosphoproteins, tryptic digestion of proteins, enrichment, and isotope labeling of phosphopeptides are performed in situ sequentially. As a result of the in situ processing design and the seamless procedures, extra steps for desalting and buffer exchanging, which are always required in conventional approaches, are avoided, and the sample loss and contamination could be greatly reduced. Thus, better sensitivity and accuracy for the quantitative phosphoproteome analysis were obtained. This strategy was further applied to differential phosphoproteome analysis of human liver tissues with or without hepatocellular carcinoma (HCC). In total, 8548 phosphorylation sites were confidently quantified from three replicate analyses of 0.5 mg of human liver protein extracts.
Language英语
WOS IDWOS:000341345000004
Citation statistics
Document Type期刊论文
Identifierhttp://cas-ir.dicp.ac.cn/handle/321008/144524
Collection中国科学院大连化学物理研究所
Corresponding AuthorYe ML(叶明亮); Zou HF(邹汉法)
Affiliation1.Chinese Acad Sci, Dalian Inst Chem Phys, Natl Chromatog R&A Ctr, CAS Key Lab Separat Sci Analyt Chem, Dalian 116023, Peoples R China
2.Univ Chinese Acad Sci, Beijing 100049, Peoples R China
3.Dalian Med Univ, Affiliated Hosp 2, Dalian 116027, Peoples R China
Recommended Citation
GB/T 7714
Huang, Junfeng,Qin, Hongqiang,Dong, Jing,et al. In Situ Sample Processing Approach (iSPA) for Comprehensive Quantitative Phosphoproteome Analysis[J]. JOURNAL OF PROTEOME RESEARCH,2014,13(9):3896-3904.
APA Huang, Junfeng.,Qin, Hongqiang.,Dong, Jing.,Song, Chunxia.,Bian, Yangyang.,...&邹汉法.(2014).In Situ Sample Processing Approach (iSPA) for Comprehensive Quantitative Phosphoproteome Analysis.JOURNAL OF PROTEOME RESEARCH,13(9),3896-3904.
MLA Huang, Junfeng,et al."In Situ Sample Processing Approach (iSPA) for Comprehensive Quantitative Phosphoproteome Analysis".JOURNAL OF PROTEOME RESEARCH 13.9(2014):3896-3904.
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