Document Detail


Use of silicone fluids in studies of cellular amino acids.
MedLine Citation:
PMID:  3086420     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
In numerous cellular studies, cells labeled with radioisotopes have been separated from the labeling medium by an aqueous solution in order to determine the quantity of internalized labels; however, the aqueous wash tends to remove significant labeling from the cells. Therefore, in order to preserve all of the internalized labels, non-aqueous medium such as silicone fluids may be used. The termination of the labeling is achieved in the silicone method when, upon centrifugation, the cells separate from the medium and enter the silicone fluid to sediment to the tube bottom. This sedimentation of cells placed above a layer of silicone fluid exhibits a critical dependence on the centrifugal force, and gives rise to an uncertainty of only 2 s in determining the time of separation of cells from the medium using General Electric F-50 silicone fluid and a modified Beckman J2-21 centrifuge. It is therefore possible to determine the kinetics of incorporation of labeled amino acids into intracellular pools and proteins. In particular, since this silicone wash method determines the size of the total pool and the aqueous wash method determines the size of the acid-extractable pool, the simultaneous measurements of the size of both pools leads to the determination of the kinetics of labeling of the free amino acid pool. Among many possible applications and extensions of these methods, the studies of formation of intracellular pools and relations among different pools of transported molecules, such as water and amino acids, appear promising.
Authors:
P N Yi; T Rhee; J O Fenn; J H Jarrett; K M Wallace
Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Journal of biochemical and biophysical methods     Volume:  12     ISSN:  0165-022X     ISO Abbreviation:  J. Biochem. Biophys. Methods     Publication Date:  1986 Mar 
Date Detail:
Created Date:  1986-06-27     Completed Date:  1986-06-27     Revised Date:  2006-11-15    
Medline Journal Info:
Nlm Unique ID:  7907378     Medline TA:  J Biochem Biophys Methods     Country:  NETHERLANDS    
Other Details:
Languages:  eng     Pagination:  123-33     Citation Subset:  IM    
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MeSH Terms
Descriptor/Qualifier:
Amino Acids / metabolism*
Animals
Biological Transport, Active
Body Fluids / metabolism*
Cell Fractionation / methods
Cells, Cultured
Centrifugation
Extracellular Space / metabolism
Intracellular Fluid / metabolism*
Kinetics
Mast-Cell Sarcoma / metabolism
Mice
Protein Biosynthesis
Silicones*
Chemical
Reg. No./Substance:
0/Amino Acids; 0/Silicones

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


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