Document Detail

Detection of apoptosis in cell-free systems.
MedLine Citation:
PMID:  19609749     Owner:  NLM     Status:  MEDLINE    
Apoptosis is a fundamental process required for proper embryonic development. Various methods have been described to detect apoptosis both in vitro as well as in vivo. Activation of caspases represents the key event in the apoptotic process. To dissect the molecular events leading to caspase activation, we have been using cell-free systems that recapitulate the mitochondrial death pathway. In the cell-free apoptosis assays, we either detect caspase activation in stimulated cells by utilizing subcellular fractions or reconstitute various components in cytosol (or mitochondria) to study molecular mechanisms of caspase activation. In either case, we utilize Western blot and/or substrate cleavage to monitor caspase activation. Using in-vitro reconstitution approach of caspase activation, we have discovered various factors that regulate caspase activity. Therefore, cell-free system not only is an invaluable tool to study apoptosis signaling but also provides molecular insight on caspase activation patterns and inhibitor specificities.
Dhyan Chandra; Dean G Tang
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Publication Detail:
Type:  Journal Article; Research Support, N.I.H., Extramural; Research Support, Non-U.S. Gov't; Research Support, U.S. Gov't, Non-P.H.S.    
Journal Detail:
Title:  Methods in molecular biology (Clifton, N.J.)     Volume:  559     ISSN:  1064-3745     ISO Abbreviation:  Methods Mol. Biol.     Publication Date:  2009  
Date Detail:
Created Date:  2009-07-17     Completed Date:  2009-10-08     Revised Date:  2010-04-29    
Medline Journal Info:
Nlm Unique ID:  9214969     Medline TA:  Methods Mol Biol     Country:  United States    
Other Details:
Languages:  eng     Pagination:  65-75     Citation Subset:  IM    
Department of Pharmacology and Therapeutics, Roswell Park Cancer Institute, Buffalo, NY, USA.
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MeSH Terms
Blotting, Western / methods
Caspases / analysis,  metabolism
Cell-Free System
Cells, Cultured
Cytochromes c / analysis,  metabolism
Cytological Techniques*
Electrophoresis, Polyacrylamide Gel / methods
Mitochondria / metabolism*
Grant Support
K017K01CA123142/CA/NCI NIH HHS; PC073751/PC/NCI NIH HHS; R01-AG023374/AG/NIA NIH HHS; R01-ES015888/ES/NIEHS NIH HHS; R21-ES015893-01A1/ES/NIEHS NIH HHS
Reg. No./Substance:
9007-43-6/Cytochromes c; EC 3.4.22.-/Caspases

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

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