Detection of an intracellular transforming protein (v-Ki-ras p21) using the flow activated cell sorter (FACS).
The transforming protein coded for by the Ki-ras oncogene, v-p21, localizes at the cytoplasmic face of the plasma membrane. A method is presented whereby the appearance of v-p21 in Kirsten murine sarcoma virus-transformed cells can be detected by flow cytometry, using a monoclonal antibody to v-p21 and methods modified from immunofluorescence microscopy. The method is sufficiently sensitive to differentiate between cellular and viral p21 levels, to detect small subpopulations of virus-transformed cells, and to monitor changes in p21 expression in response to physiologic variables. The method provides a rapid, quantitative means to investigate the expression of an intracellular transforming protein in heterogeneous cell populations.
D000818:Animals; D002472:Cell Transformation, Viral; D002478:Cells, Cultured; D005260:Female; D005434:Flow Cytometry; D006107:Granulosa Cells; D007668:Kidney; D007708:Kirsten murine sarcoma virus; D009363:Neoplasm Proteins / Q000032:analysis*; D015689:Oncogene Protein p21(ras); D051381:Rats; D012513:Sarcoma, Experimental / Q000032:analysis*
D009363:Neoplasm Proteins; D015689:Oncogene Protein p21(ras)