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 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.
Appel
M
M
Couderc
E
E
Feger
J
J
D000418:Albumins / Q000096:biosynthesis; D000818:Animals; D002452:Cell Count; D002478:Cells, Cultured; D003921:Diabetes Mellitus, Experimental / Q000378:metabolism; D007553:Isotope Labeling / Q000592:standards*; D008099:Liver / Q000378:metabolism; D008297:Male; D008715:Methionine; D008722:Methods; D014176:Protein Biosynthesis; D051381:Rats; D011919:Rats, Inbred Strains; D013462:Sulfur Radioisotopes
D000418:Albumins; D013462:Sulfur Radioisotopes; D008715:Methionine