Selected article for: "chase pulse labeling and pulse labeling"
Author: Maceyka, Michael; Machamer, Carolyn E.
Title: Ceramide Accumulation Uncovers a Cycling Pathway for the cis-Golgi Network Marker, Infectious Bronchitis Virus M Protein
  • Document date: 1997_12_15
    • ID: wekvet6f_10
    Snippet: Pulse-Chase Labeling. Pulse-chase experiments were performed as previously described (Rosenwald et al., 1992; Weisz et al., 1993) . Briefly, cells were plated in 35-mm dishes the night before the experiment to be 90% confluent the next day. VSV (San Juan strain, Indiana serotype) was adsorbed for 30 min in 0.5 ml of serum-free DME. At 4 h after infection, the cells were starved for methionine for 15 min. Cells were then pulsed for 5 min with 50 C.....
    Document: Pulse-Chase Labeling. Pulse-chase experiments were performed as previously described (Rosenwald et al., 1992; Weisz et al., 1993) . Briefly, cells were plated in 35-mm dishes the night before the experiment to be 90% confluent the next day. VSV (San Juan strain, Indiana serotype) was adsorbed for 30 min in 0.5 ml of serum-free DME. At 4 h after infection, the cells were starved for methionine for 15 min. Cells were then pulsed for 5 min with 50 Ci 35 S-Pro-mix and chased for the indicated amount of time in the presence or absence of the indicated drugs. The isopropanol carrier alone had no effect on the rate of transport of VSV G protein. After the chase, cells were washed once with cold PBS and lysed in detergent. VSV G protein was then immunoprecipitated, treated with endo H as described (Rosenwald et al., 1992) , separated by SDS-PAGE, and visu-alized by fluorography. Endo H-sensitive and -resistant forms of the VSV G protein were quantitated by densitometry.

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