Document Detail

A method to measure receptor binding of ligands with low affinity. Application to plasma proteins binding assay with hemopoietic cells.
MedLine Citation:
PMID:  2981695     Owner:  NLM     Status:  MEDLINE    
A gradient has been developed for separating free ligand from ligand bound to cells growing in suspension. This method can be used with all kinds of ligand but it is particularly useful for those ligands having the tiresome tendency to adhere to the cells non-specifically or to polymerize by themselves. This is the case of fibronectin, fibrinogen, immunoglobulins and many other plasma proteins. The gradient consists of two layers: an upper aqueous phase and a lower hydrophobic organic phase. The aqueous phase, a sucrose buffer, allows the cells to become well dispersed before they enter the hydrophobic phase which excludes the free ligand efficiently. This reduces non-specific binding and allows the accurate measurement of specific binding which could not be obtained with a gradient made of a single phase. Depending upon the size and the density of the cells, and the nature of the ligand, the assay method can be modified by changing the density or the nature of the hydrophilic and hydrophobic phases.
J P Levesque; A Hatzfeld; J Hatzfeld
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Experimental cell research     Volume:  156     ISSN:  0014-4827     ISO Abbreviation:  Exp. Cell Res.     Publication Date:  1985 Feb 
Date Detail:
Created Date:  1985-03-20     Completed Date:  1985-03-20     Revised Date:  2006-11-15    
Medline Journal Info:
Nlm Unique ID:  0373226     Medline TA:  Exp Cell Res     Country:  UNITED STATES    
Other Details:
Languages:  eng     Pagination:  558-62     Citation Subset:  IM    
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MeSH Terms
Cell Count
Cell Line
Centrifugation, Density Gradient
Dibutyl Phthalate
Fibrinogen / metabolism*
Lymphocytes / metabolism*
Phthalic Acids
Platelet Membrane Glycoproteins
Receptors, Cell Surface / metabolism*
Reg. No./Substance:
0/Phthalic Acids; 0/Platelet Membrane Glycoproteins; 0/Receptors, Cell Surface; 131-11-3/dimethyl phthalate; 84-74-2/Dibutyl Phthalate; 9001-32-5/Fibrinogen

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

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