Simultaneous quantification of c-myc oncoprotein, total cellular protein, and DNA content using multiparameter flow cytometry.

摘要: Variations in total cellular protein content can confound interpretation of the significance modulations specific proteins. In an effort to overcome this problem, a technique is described for simultaneous measurement protein, and DNA content. The method utilizes dual-laser (uv 488 nm) excitation three fluorescent dyes: FITC, SR101, DAPI. FITC-labelled antibody coupled with indirect immunofluorescence was used quantify c-myc oncoprotein, whereas SR101 DAPI were measure DNA, respectively. Flow cytometric measurements oncoprotein compared densitometric readings p64c-myc. determinations those obtained by Lowry technique. Results indicated that flow correlated well biochemical methods. usefulness further examined following treatment exponentially growing HL-60 cells 2.5 micrograms/ml cycloheximide 0 12 h. Cycloheximide found cause significant decrease within 2 h (P less than 0.05), relative increase proportion G0/G1 modest protein. This appears provide rapid, quantitative approach, useful investigating alterations growth balance occurring cell differentiation, neoplastic transformation, or radiation cytostatic drugs.