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Comparative studies of protein biosynthesis: the main experimental parameters in pulse and pulse‐chase experiments must be standardized
Author(s) -
Appel Martine,
Couderc Elisabeth,
Feger Jeanne
Publication year - 1992
Publication title -
biology of the cell
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.543
H-Index - 85
eISSN - 1768-322X
pISSN - 0248-4900
DOI - 10.1016/0248-4900(92)90031-u
Subject(s) - biology , methionine , pulse (music) , intracellular , compartment (ship) , hepatocyte , amino acid , function (biology) , protein biosynthesis , biochemistry , biophysics , microbiology and biotechnology , in vitro , physics , oceanography , detector , optics , geology
Summary— The influence of experimental conditions was investigated in pulse and pulse‐chase experiments involving l ‐[ 35 s]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.