Pulse-chase labeling

In Situ Generation of Coherently Labeled Precursor Populations - Analyzing the Biosynthesis of Plant Metabolites via Photosynthetic Pulse/Chase Labeling with... 

The biosynthesis of arylsulfohydrolase B has been studied in hamster and human skin fibroblasts (Warburton and Wynn, 1976 Steckel et al., 1983). Pulse-chase labeling and uptake studies have indicated that arylsulfohydrolase B is synthesized and secreted as a 64,000 precursor. This precursor is... 

Pulse-chase labeling can determine the Intracellular fate of proteins and other metabolites (see Figure 3-36). 

Transport of newly synthesized PC from the ER to the plasma membrane. The principal site of PC synthesis is the ER and Golgi (Chapter 8). The transport of PC from its sites of synthesis to the plasma membrane has been examined using pulse-chase labeling with a [ Hjcholine precursor for PC, and rapid plasma membrane isolation with cationic beads (M. Kaplan, 1985). These studies reveal that PC transport is an extremely rapid process occurring with a 1 min (Fig. 8). This transport is unaffected by metabolic poisons that deplete cellular ATP levels, disrupt vesicle transport, or alter cytoskeletal arrangement. 

The Biarsenical-tetracysteine Protein Tag Chemistry and Biological Applications 443 8.1.4.2 Multicolor Pulse-chase Labeling... 

The evidence for this mode of chain extension comes chiefly from the studies of Robbins etal (1967), who used pulse-chase labelling to show that a mutant of S. typhimurium that could not synthesise a complete core structure would assemble oligosaccharide chains in which the most recently inserted residues were near the reducing end. A similar mechanism could be shown both in cell-free particulate systems and the intact organisms. The individual steps in this process have not been separated in vitro and the intermediates with longer saccharide chains cannot readily be isolated without degradation. 

The first protocol is the classical procedure to visualize nascent replicating DNA by fluorescence microscopy. This method can be combined with other immunolabeling methods, although it should be kept in mind that the rather harsh DNA denaturation conditions may interfere with other labeling procedures. The protocol has the advantage that replicating DNA can be pulse-chase labeled, because incorporation of halo-dU nucleotides takes place in vivo and BrdU is rather nontoxic (however, see Stone and Stephens, 1993). 

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