Document Detail


Centrifugal elutriation to obtain synchronous populations of cells.
MedLine Citation:
PMID:  18228390     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Counterflow centrifugal elutriation is a noninvasive method for separating large numbers of cells on the basis of their size and mass. For mammalian cells, this method is useful for separating mixed populations of cells, in particular cells at different stages of the cell division cycle without perturbing cell metabolism or using synchronizing agents. This unit describes a method for separating 2 x 10(8) cells using the standard JE-6B rotor or larger numbers of cells in the JE-5.0 rotor. To verify the purity and to characterize the cell cycle positions of cells in the elutriated populations, the unit includes protocols for measuring nascent DNA synthesis by [(3)H]thymidine incorporation and for detecting DNA synthesis and content by propidium iodide flow cytometry alone or in combination with bromodeoxyuridine incorporation.
Authors:
A F Wahl; K L Donaldson
Publication Detail:
Type:  Journal Article    
Journal Detail:
Title:  Current protocols in cell biology / editorial board, Juan S. Bonifacino ... [et al.]     Volume:  Chapter 8     ISSN:  1934-2616     ISO Abbreviation:  Curr Protoc Cell Biol     Publication Date:  2001 May 
Date Detail:
Created Date:  2008-01-29     Completed Date:  2008-02-20     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 8.5     Citation Subset:  IM    
Affiliation:
Seattle Genetics, Inc., Bothell, Washington, USA.
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MeSH Terms
Descriptor/Qualifier:
Animals
Bromodeoxyuridine / analysis
Cell Line, Tumor / cytology
Cell Separation / instrumentation,  methods*
Centrifugation / instrumentation,  methods*
DNA / analysis
DNA Replication
Flow Cytometry / methods
Fluorescent Dyes / analysis
Humans
Propidium / analysis
Staining and Labeling
Thymidine / metabolism
Tritium / analysis
Chemical
Reg. No./Substance:
0/Fluorescent Dyes; 10028-17-8/Tritium; 36015-30-2/Propidium; 50-89-5/Thymidine; 59-14-3/Bromodeoxyuridine; 9007-49-2/DNA

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


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