Document Detail


Isolation of peroxisomes from tissues and cells by differential and density gradient centrifugation.
MedLine Citation:
PMID:  18228357     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Peroxisome purification depends on a two-step procedure: differential centrifugation to prepare a light mitochondrial fraction and fractionation on a density-gradient medium preferably iodixanol or Nycodenz, to isolate the peroxisome enriched fraction. The iodixanol gradient may be a preformed continuous gradient or a self-generating gradient. Alternatively a continuous Nycodenz gradient or a simple Nycodenz barrier may be used for the second step. The unit contains protocols for peroxisome isolation from rat liver, tissue culture cells (HepG2 cells), and yeast spheroplasts. The extent of endoplasmic reticulum contamination of the prep can be assessed using an assay for the marker enzyme NADPH-cytochrome creductase.
Authors:
J M Graham
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Publication Detail:
Type:  Journal Article    
Journal Detail:
Title:  Current protocols in cell biology / editorial board, Juan S. Bonifacino ... [et al.]     Volume:  Chapter 3     ISSN:  1934-2616     ISO Abbreviation:  Curr Protoc Cell Biol     Publication Date:  2001 May 
Date Detail:
Created Date:  2008-01-29     Completed Date:  2008-03-24     Revised Date:  2009-12-11    
Medline Journal Info:
Nlm Unique ID:  101287856     Medline TA:  Curr Protoc Cell Biol     Country:  United States    
Other Details:
Languages:  eng     Pagination:  Unit 3.5     Citation Subset:  IM    
Affiliation:
Liverpool John Moores University, Liverpool, United Kingdom.
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MeSH Terms
Descriptor/Qualifier:
Animals
Biological Markers
Carcinoma, Hepatocellular / pathology
Cell Fractionation / methods*
Cell Line, Tumor / ultrastructure
Centrifugation / methods*
Centrifugation, Density Gradient / methods
Endoplasmic Reticulum / enzymology
Humans
Iohexol
Liver / ultrastructure
Liver Neoplasms / pathology
NADPH-Ferrihemoprotein Reductase / analysis
Peroxisomes*
Rats
Rats, Sprague-Dawley
Saccharomyces cerevisiae / ultrastructure
Spheroplasts / ultrastructure
Triiodobenzoic Acids
Chemical
Reg. No./Substance:
0/Biological Markers; 0/Triiodobenzoic Acids; 66108-95-0/Iohexol; 92339-11-2/iodixanol; EC 1.6.2.4/NADPH-Ferrihemoprotein Reductase

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


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