Document Detail


A modified tandem affinity purification tag technique for the purification of protein complexes in mammalian cells.
MedLine Citation:
PMID:  15602769     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
The tandem affinity purification (TAP) tag technique has been used with success to identify under nondenaturing conditions protein complexes in yeast. The technique can be used in mammalian cells, but we found that the original technique does not yield enough recovery for the identification of proteins when mammalian cells growing in monolayer have to be used. We present here a modified TAP tag technique that allows sufficient recovery of proteins from mouse fibroblasts growing in monolayer cultures. The recovery allows protein identification by mass spectrometry.
Authors:
Robert Drakas; Marco Prisco; Renato Baserga
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Publication Detail:
Type:  Journal Article; Research Support, N.I.H., Extramural; Research Support, U.S. Gov't, P.H.S.    
Journal Detail:
Title:  Proteomics     Volume:  5     ISSN:  1615-9853     ISO Abbreviation:  Proteomics     Publication Date:  2005 Jan 
Date Detail:
Created Date:  2005-02-24     Completed Date:  2005-08-08     Revised Date:  2008-11-21    
Medline Journal Info:
Nlm Unique ID:  101092707     Medline TA:  Proteomics     Country:  Germany    
Other Details:
Languages:  eng     Pagination:  132-7     Citation Subset:  IM    
Affiliation:
Kimmel Cancer Center, Thomas Jefferson University, Philadelphia, PA 19107, USA.
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MeSH Terms
Descriptor/Qualifier:
Affinity Labels / chemistry*
Animals
Biotinylation
Cell Line
Cell Proliferation
Embryo, Mammalian / cytology
Escherichia coli Proteins / genetics
Fibroblasts / chemistry,  cytology
Humans
Immunoprecipitation
Insulin Receptor Substrate Proteins
Insulin-Like Growth Factor I / pharmacology
Mass Spectrometry
Mice
Multiprotein Complexes / chemistry,  genetics,  isolation & purification*
Phosphoproteins / chemistry,  genetics,  isolation & purification*
Plasmids
Recombinant Fusion Proteins / chemistry,  genetics,  isolation & purification
Grant Support
ID/Acronym/Agency:
AG20956/AG/NIA NIH HHS; CA86940/CA/NCI NIH HHS
Chemical
Reg. No./Substance:
0/Affinity Labels; 0/Escherichia coli Proteins; 0/IRS1 protein, human; 0/Insulin Receptor Substrate Proteins; 0/Irs1 protein, mouse; 0/Multiprotein Complexes; 0/Phosphoproteins; 0/Recombinant Fusion Proteins; 67763-96-6/Insulin-Like Growth Factor I

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


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