A method for sequential quantitative separation and determination of protein, RNA, DNA, lipid, and glycogen from a single rat liver homogenate or from a subcellular fraction

✍ Scribed by S. Shibko; P. Koivistoinen; C.A. Tratnyek; A.R. Newhall; L. Friedman

• Publisher: Elsevier Science
• Year: 1967
• Tongue: English
• Weight: 828 KB
• Volume: 19
• Category: Article
• ISSN: 0003-2697
• DOI: 10.1016/0003-2697(67)90242-4

✦ Synopsis

An accepted method for studying the metabolic activity of the cell under experimental conditions has been to measure its protein, RNA and DNA composition, and the incorporation of specific precursors into these components. This gives valuable information as to the functioning of the mechanisms involved in protein synthesis. In many cases the process has been extended to study these components in subcellular fractions. In order to evaluate the function of all the major metabolic areas, however, it is desirable to extend these studies to include other major cellular constituents such as glycogen and lipid. These measurements should be carried out simultaneously on the principal subcellular fractions of the cell. Further, if suitably labeled substances known to be involved in many metabolic pathways are administered to the animal before sacrifice, it should be possible to detect changes in the rates of metabolism as well as in the composition of these major components. Such measurements should provide clues as to the sites of action during an experimental treatment that would guide more detailed studies of the mechanisms involved in the production of observed effects.

In order to measure the incorporation of radioactivity into these specific substances, a scheme for chemical fractionation of the components of the cell is required which allows "quantitative" recovery of each component with a minimum of cross contamination of the other components. The method should be applicable to all the subcellular fractions 'Contribution No. 1009 from the Department of Nutrition and Food Science,