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Immunofluorescence assays. For MDV immunofluorescence assays,
CEFs were plated on poly-L-lysine coated 12 mm diameter glass dishes
(12-well plates) at 8 ¡¤ 104 cells well/ml density in 10% FCS medium.
The following day, medium was removed and MDV was inoculated at
m.o.i. of 1 PFU/cell, in 2% FCS MEM, in the presence and absence of
proteins. After 5 h of incubation at 37 C in 5%CO2, monolayers were
washed twice with PBS and then fixed with 4% p-formaldehyde in PBS
for 30 min at room temperature. After washing twice more, 0.1M
glycine was added to neutralize eventual residual traces of fixative.
Fixed cells were left overnight at 4 C in PBS. For intracellular
detection of MDV-specific signals by fluorescence microscopy, cells
were permeabilized with 0.1% Triton X-100 for 5 min to allow antibodies
to enter the cytoplasm and interact with viral antigens. Fluorescein
conjugated (FITC) anti-MDV chicken antibody was purchased
from Eurobio (France).
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Сµ¤Ä¾Ä¾(½ð±Ò+3, MolEPI+1): ºÜÈÈÐÄŶ 2012-02-28 16:32:46
ÊÍÃÎ110(½ð±Ò+20): ¡ïÓаïÖú ËäÈ»ÎÒÒѾ­·­Òë³öÀ´ÁË£¬µ«»¹ÊǷdz£·Ç³£µÄ¸Ðл£¡ 2012-02-29 12:52:18
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