Document Detail


Antibodies immobilized as arrays to profile protein post-translational modifications in mammalian cells.
MedLine Citation:
PMID:  15123764     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Previously, we demonstrated that antibodies printed on a solid support were able to detect protein-protein interaction in mammalian cells. Here we further developed the antibody array system for detecting proteins with various post-translational modifications in mammalian cells. In this novel approach, immunoprecipitated proteins were labeled with fluorescent dye followed by incubation over antibody arrays. Targeted proteins, captured by the antibodies immobilized on PVDF membrane or glass slide, were detected by means of near infrared fluorescent scanner or fluorescent microscopy. To demonstrate the application of the antibody arrays in protein post-translational modifications, we profiled protein tyrosine phosphorylation, ubiquitination, and acetylation in mammalian cells under different conditions. Our results indicate that antibody array technology can provide a powerful means of profiling a large number of proteins with different post-translational modifications in cells.
Authors:
Stanimir S Ivanov; Alicia S Chung; Zheng-long Yuan; Ying-jie Guan; Katherine V Sachs; Jonathan S Reichner; Y Eugene Chin
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Publication Detail:
Type:  Journal Article; Research Support, U.S. Gov't, P.H.S.     Date:  2004-05-03
Journal Detail:
Title:  Molecular & cellular proteomics : MCP     Volume:  3     ISSN:  1535-9476     ISO Abbreviation:  Mol. Cell Proteomics     Publication Date:  2004 Aug 
Date Detail:
Created Date:  2004-08-16     Completed Date:  2005-03-11     Revised Date:  2007-11-14    
Medline Journal Info:
Nlm Unique ID:  101125647     Medline TA:  Mol Cell Proteomics     Country:  United States    
Other Details:
Languages:  eng     Pagination:  788-95     Citation Subset:  IM    
Affiliation:
Department of Pathology & Laboratory Medicine, Brown University Medical School, Providence, RI 02912, USA.
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MeSH Terms
Descriptor/Qualifier:
3T3 Cells
Acetylation
Animals
Antibodies*
Cell Line
Fluorescent Antibody Technique
Fluorescent Dyes
Gene Expression Profiling
Hela Cells
Humans
Mice
Phosphorylation
Protein Array Analysis*
Protein Processing, Post-Translational*
Tyrosine / metabolism*
Ubiquitin / metabolism*
Grant Support
ID/Acronym/Agency:
GM-66194/GM/NIGMS NIH HHS; R01 CA-82549/CA/NCI NIH HHS
Chemical
Reg. No./Substance:
0/Antibodies; 0/Fluorescent Dyes; 0/Ubiquitin; 55520-40-6/Tyrosine

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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