PRODUCTION OF MONOCLONAL-ANTIBODIES REACTIVE WITH A DENATURED FORM OF THE FRIEND MURINE LEUKEMIA-VIRUS GP70 ENVELOPE PROTEIN - USE IN A FOCAL INFECTIVITY ASSAY, IMMUNOHISTOCHEMICAL STUDIES, ELECTRON-MICROSCOPY AND WESTERN BLOTTING

被引:111
作者
ROBERTSON, MN
MIYAZAWA, M
MORI, S
CAUGHEY, B
EVANS, LH
HAYES, SF
CHESEBRO, B
机构
[1] NIAID,PERSISTENT VIRAL DIS LAB,ROCKY MT LABS,903 S 4TH ST,HAMILTON,MT 59840
[2] NIAID,VECTORS & PATHOGENS LAB,ROCKY MT LABS,HAMILTON,MT 59840
关键词
MONOCLONAL ANTIBODY; RETROVIRUS; IMMUNOHISTOCHEMISTRY; ELECTRON MICROSCOPY;
D O I
10.1016/0166-0934(91)90105-9
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
Four monoclonal antibodies were selected for their ability to recognize the envelope protein of Friend murine leukemia virus (F-MuLV) in methanol-fixed tissue culture cells. Each of these monoclonal antibodies was found to react only with F-MuLV. By using recombinant retroviruses, it was determined that each of the monoclonal antibodies recognized the C-terminal one-third of the F-MuLV gp70 envelope protein. The monoclonal antibodies were effective in radioimmunoprecipitation of F-MuLV proteins, and one of the antibodies, 720, was also effective in Western blotting. The ability of antibody 720 to react with F-MuLV in methanol-fixed cells facilitated the use of a sensitive immunoperoxidase method with a focal virus infectivity assay. In immunohistochemical studies using light microscopy, antibody 720 could specifically label F-MuLV-infected cells in acetone-fixed tissue sections from F-MuLV-infected animals. Finally, in immuno-gold labelling studies using electron microscopy, antibody 720 could be used to distinguish F-MuLV from amphotropic MuLV.
引用
收藏
页码:255 / 271
页数:17
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