Localization of Epstein-Barr Virus Early Antigen (EA) by Electron Microscopy
The localization of polypeptides (50–52,85K) associated with the diffuse and restricted components of early antigen EA in Epstein-Barr virus (EB virus)- infected cells was demonstrated with immune electron microscopy (EM) using colloidal — gold staining. P3HR-1 cells were activated to express the viral antigens by treatment with 4 mM n-butyrate and 20 ng/ml TPA. After 32–38 hrs, the cells were fixed with 2% glutaraldehyde, dehydrated with alcohol and propylene oxide, and embeded in epon. Ultra-thin section were prepared and incubated first with the appropriate monoclonal antibody, then with Staphylococcal protein A — gold, and finally with uranyl acetate. EM examination revealed that the 50–52 K component of EA-D was localized both in nucleoplasm and cytoplasm of EA-positive cells, while the 85 K component of EA-R was located mainly in the cytoplasm, as indicated by specific colloidal-gold staining.
KeywordsPropylene Oxide Osmic Acid Early Antigen Staphylococcal Protein Immune Electron Microscopy
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