Author: Lam, Yun W.; Evans, Vanessa C.; Heesom, Kate J.; Lamond, Angus I.; Matthews, David A.
Title: Proteomics Analysis of the Nucleolus in Adenovirus-infected Cells Document date: 2009_10_7
ID: jgxbpy4j_15
Snippet: Antibodies and Plasmids-Antisera used were anti-adenovirus protein V ( Confocal Microscopy-HeLa cells grown on coverslips were either infected, mock infected, and/or transfected with expression plasmids encoding tagged proteins as indicated. At 18 h postinfection, cells were fixed for 5 min with 4% formaldehyde in PBS followed by washing in PBS before permeabilizing in 1% Triton X-100 in PBS for 5 min. After permeabilization, the cells were washe.....
Document: Antibodies and Plasmids-Antisera used were anti-adenovirus protein V ( Confocal Microscopy-HeLa cells grown on coverslips were either infected, mock infected, and/or transfected with expression plasmids encoding tagged proteins as indicated. At 18 h postinfection, cells were fixed for 5 min with 4% formaldehyde in PBS followed by washing in PBS before permeabilizing in 1% Triton X-100 in PBS for 5 min. After permeabilization, the cells were washed briefly with PBS before blocking for 30 min at room temperature in 10% FCS in PBS. The cells were then incubated with primary antibodies for 60 min, washed, and incubated with secondary antibodies. Cells were mounted with Vectashield plus DAPI to visualize the nuclear DNA. Images were taken using a Leica confocal microscope (TCS-SP2).
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