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_11
Snippet: Indirect Immunofluorescence Microscopy. These experiments were performed as previously described (Swift and Machamer, 1991) . Briefly, cells were infected for 30 min with a recombinant vaccinia virus encoding IBV M, and the indicated treatments were begun 4 h after infection. For experiments using exogenous ceramides, the soluble short-chain analogues of ceramide were added to cells as a complex with 0.34 mg/ml defatted BSA as described (Pagano a.....
Document: Indirect Immunofluorescence Microscopy. These experiments were performed as previously described (Swift and Machamer, 1991) . Briefly, cells were infected for 30 min with a recombinant vaccinia virus encoding IBV M, and the indicated treatments were begun 4 h after infection. For experiments using exogenous ceramides, the soluble short-chain analogues of ceramide were added to cells as a complex with 0.34 mg/ml defatted BSA as described (Pagano and Martin, 1988) . After treatment, cells were fixed, permeabilized, and stained with the appropriate antibody. Images were acquired using a microscope (Axioskop; Zeiss, Inc., Thornwood, NY) equipped with epifluorescence and a CCD camera (Photometrics Sensys, Tucson, AZ) using IP Lab software (Signal Analytics Corp., Vienna, VA). All images shown are the raw data collected at 1 Ï« 1 binning with a gain of 1.
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