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SMALL-SCALE ISOLATION OF FERRITIN FOR THE ASSAY OF THE INCORPORATION OF 14C-LABELLED AMINO ACIDS.

Authors
  • DRYSDALE, J W
  • MUNRO, H N
Type
Published Article
Journal
The Biochemical journal
Publication Date
Jun 01, 1965
Volume
95
Pages
851–858
Identifiers
PMID: 14342525
Source
Medline
Keywords
License
Unknown

Abstract

1. A new procedure is described for the isolation of pure ferritin from small amounts of tissue. After the removal of most of the tissue proteins by heat coagulation, the ferritin fraction was chromatographed successively on CM-cellulose and Sephadex G-200. 2. The isolated ferritin appeared to be free from other proteins, as judged by its sedimentation pattern in the ultracentrifuge, by electrophoresis on polyacrylamide gel and by immunoelectrophoresis. 3. After the injection of [(14)C]leucine into a series of rats, the specific activity of liver ferritin isolated by the new procedure bore a constant relationship to that of liver ferritin separated by antigen-antibody precipitation. The procedure can thus be used to obtain ferritin of suitable purity for studies of amino acid incorporation. 4. The new procedure can be used to measure the total ferritin protein content of a tissue. It is not possible to use ultraviolet absorption for the measurement of ferritin protein because of considerable interference from the iron that it contains.

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