Molecular quantification of cell cycle‐related gene expression at the protein level

摘要: BACKGROUND Immunofluorescence cytometry of antigen and DNA content provides relative measurements the cell cycle phase distribution a specific epitope. Measurement correlated expression epitopes on signaling regulatory proteins will be useful in study complex pathways involved regulation carcinogenesis. However, to formulate pathway models, molecules per would more than intensity. Here, we report system which relationship between fluorescence is determined for reference set lines that are then used directly calculate number unknowns. To demonstrate process, calculated SV40 large T (Tag) cells. METHODS A line clones expressing different levels Tag were isolated. Quantitative Western blots these cells purified, recombinant performed. Cells from same sample stained analyzed by flow DNA. The was established calculations performed distributions Tag. RESULTS The five had 0.11, 0.27, 1.06, 2.44, 2.63 × 106 cell, blot. average coefficient variation 10.6%. fit linear equation (r2 = 0.96) over range, 0.11 – molecules, however, did not 0 defined isotype staining controls, lowest line. line, second 110,000 used. CONCLUSIONS This work describes where fixed various target quantified can standardize cytometric gene absolute terms.Cytometry 39:79–89, 2000 © Wiley-Liss, Inc.