Comparative studies of protein biosynthesis: the main experimental parameters in pulse and pulse-chase experiments must be standardized

The influence of experimental conditions was investigated in pulse and pulse-chase experiments involving l-[ 35s]methionine incorporation by isolated rat hepatocytes. The incorporation of the labelled amino acid must be linear as a function of dose and time, and similar hepatocyte densities should b...

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Bibliographic Details
Published in:Biology of the cell 1992, Vol.74 (2), p.235-238
Main Authors: Appel, Martine, Couderc, Elisabeth, Feger, Jeanne
Format: Article
Language:English
Subjects:
Albumins - biosynthesis
Animals
Cell Count
Cells, Cultured
chase
Diabetes Mellitus, Experimental - metabolism
hepatocyte
Isotope Labeling - standards
Liver - metabolism
Male
Methionine
Methods
Protein Biosynthesis
pulse
Rats
Rats, Inbred Strains
Sulfur Radioisotopes
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Summary:The influence of experimental conditions was investigated in pulse and pulse-chase experiments involving l-[ 35s]methionine incorporation by isolated rat hepatocytes. The incorporation of the labelled amino acid must be linear as a function of dose and time, and similar hepatocyte densities should be used to compare radioactive uptake. High hepatocyte densities give greater precision. In pulse-chase experiments, it was shown that a pool of free radioactivity moves progressively during the chase phase from the intracellular compartment. Protein-associated radioactivity must therefore be expressed as a function of both total radioactive uptake and total labelled protein.
ISSN:0248-4900
1768-322X
DOI:10.1016/0248-4900(92)90031-U