Document Detail


Combining microinjection and immunoblotting to analyze MAP kinase phosphorylation in single starfish oocytes and eggs.
MedLine Citation:
PMID:  19085132     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
The starfish oocyte has proven useful for studies involving microinjection because it is relatively large (190 mum) and optically clear. These oocytes are easily obtained from the ovary arrested at prophase of meiosis I, making them useful as a model system for the study of cell cycle-related events. In this chapter, a method for combining microinjection with immunoblotting of single cells is described. Individual starfish oocytes are injected, removed from the microinjection chamber, and analyzed by immunoblotting for the dual-phosphorylated form of mitogen-activated protein kinase (MAPK). This method will allow for experiments testing the regulation of MAPK in single cells and for the manipulation of these cells by a quantitative microinjection technique.
Authors:
David J Carroll; Wei Hua
Publication Detail:
Type:  Journal Article    
Journal Detail:
Title:  Methods in molecular biology (Clifton, N.J.)     Volume:  518     ISSN:  1064-3745     ISO Abbreviation:  Methods Mol. Biol.     Publication Date:  2009  
Date Detail:
Created Date:  2008-12-16     Completed Date:  2009-02-11     Revised Date:  2009-11-19    
Medline Journal Info:
Nlm Unique ID:  9214969     Medline TA:  Methods Mol Biol     Country:  United States    
Other Details:
Languages:  eng     Pagination:  57-66     Citation Subset:  IM    
Affiliation:
Department of Biological Sciences, Florida Institute of Technology, Melbourne, FL, USA.
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MeSH Terms
Descriptor/Qualifier:
Animals
Dissection
Electrophoresis, Agar Gel
Immunoblotting / methods*
Microinjections / methods*
Mitogen-Activated Protein Kinases / metabolism*
Oocytes / enzymology*
Phosphorylation
Starfish / cytology*,  enzymology*
Chemical
Reg. No./Substance:
EC 2.7.11.24/Mitogen-Activated Protein Kinases

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


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