Quantitative analysis of a nuclear antigen in interphase and mitotic cells.
The quantification of an interchromatin-associated antigen, designated p 105, during cellular passage through mitosis is described. Indirect immunofluorescence microscopy and immunogold electron microscopy demonstrated a qualitative increase in p 105 within the mitotic cytoplasm. Multiparameter flow cytometric analysis was performed on fixed cells sequentially stained with anti-p 105 immunofluorescence and/or propidium iodide. This analysis demonstrated approximately a tenfold increase in intracellular p 105 content as a function of progression from the G2 to the M phase. This increase was corroborated by the quantitative immunoblot analysis of colchicine-treated cell cultures and of cells sorted on the basis of anti-p 105 immunofluorescence. The data reveal that the increased levels of anti-p 105 immunofluorescence in conjunction with flow cytometry may be used effectively to quantitate mitotic index and isolate mitotic cells. The function and modulation of p 105 throughout the cell cycle is discussed.
Clevenger
C V
CV
Epstein
A L
AL
Bauer
K D
KD
D000936:Antigen-Antibody Complex; D000941:Antigens / Q000032:analysis; D002460:Cell Line; D002467:Cell Nucleus / Q000648:ultrastructure*; D002843:Chromatin / Q000648:ultrastructure*; D005434:Flow Cytometry / Q000379:methods; D006367:HeLa Cells / Q000166:cytology; D006801:Humans; D007124:Immunoenzyme Techniques; D007399:Interphase; D008854:Microscopy, Electron / Q000379:methods; D008938:Mitosis; D008940:Mitotic Index; D009698:Nucleoproteins / Q000032:analysis*
D000936:Antigen-Antibody Complex; D000941:Antigens; D002843:Chromatin; D009698:Nucleoproteins