Metabolic labeling with amino acids.
Metabolic labeling techniques are used to study the biosynthesis, processing, intracellular transport, secretion, degradation, and physical-chemical properties of proteins. This unit provides protocols for metabolic labeling of cells with [(35)S]-labeled amino acids to provide material suitable for analysis by immunoprecipitation, for characterization of cellular proteins, for analysis of protein trafficking, and for one- and two-dimensional gel electrophoresis. Three procedures are described for suspension and adherent cells: pulse-labeling, pulse-chase labeling, or continuous long-term labeling. A support protocol describes TCA precipitation to measure the extent of labeling.
Bonifacino
J S
JS
National Institute of Child Health and Human Development, Bethesda, Maryland, USA.
D000596:Amino Acids / Q000032:analysis* / Q000378:metabolism; D000818:Animals; D002478:Cells, Cultured / Q000378:metabolism; D011232:Chemical Precipitation; D007553:Isotope Labeling / Q000379:methods*; D008322:Mammals / Q000378:metabolism; D008715:Methionine / Q000032:analysis / Q000378:metabolism; D011506:Proteins / Q000032:analysis* / Q000302:isolation & purification / Q000378:metabolism; D013462:Sulfur Radioisotopes / Q000032:analysis / Q000378:metabolism; D014238:Trichloroacetic Acid
D000596:Amino Acids; D011506:Proteins; D013462:Sulfur Radioisotopes; D014238:Trichloroacetic Acid; D008715:Methionine