用于检测¹⁴C标记氨基酸掺入情况的铁蛋白小规模分离法。
SMALL-SCALE ISOLATION OF FERRITIN FOR THE ASSAY OF THE INCORPORATION OF 14C-LABELLED AMINO ACIDS.
作者信息
DRYSDALE J W, MUNRO H N
出版信息
Biochem J. 1965 Jun;95(3):851-8. doi: 10.1042/bj0950851.
Abstract
- 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.
摘要
- 本文描述了一种从小量组织中分离纯铁蛋白的新方法。通过热凝固去除大部分组织蛋白后，铁蛋白组分先后在CM-纤维素和葡聚糖凝胶G-200上进行层析。2. 通过超速离心的沉降模式、聚丙烯酰胺凝胶电泳和免疫电泳判断，分离出的铁蛋白似乎不含其他蛋白质。3. 给一系列大鼠注射[(14)C]亮氨酸后，用新方法分离的肝脏铁蛋白的比活性与通过抗原-抗体沉淀分离的肝脏铁蛋白的比活性保持恒定关系。因此，该方法可用于获得适合研究氨基酸掺入的纯度合适的铁蛋白。4. 新方法可用于测量组织中铁蛋白的总蛋白含量。由于铁蛋白所含铁的显著干扰，无法用紫外吸收法测量铁蛋白蛋白。
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本文引用的文献
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