Cham B E, Roeser H P, Nikles A, Ridgway K
Anal Biochem. 1985 Dec;151(2):561-5. doi: 10.1016/0003-2697(85)90220-9.
A simple, rapid technique for purification of ferritin from human liver tissue is described. Methanol, at a final concentration of 40% (v/v) in liver homogenate, precipitates the majority of proteins but does not affect ferritin. Subsequent heating of this homogenate at 75 degrees C for 10 min results in a purified ferritin preparation as judged by immunoelectrophoresis and polyacrylamide gel electrophoresis. The resultant purified ferritin contained the same amount of iron as the original endogenous ferritin. There were no significant differences (paired t tests) in the amount of protein in the purified ferritin preparation when measured by rocket immunoelectrophoresis and by the Lowry procedure, suggesting that the antigenecity of ferritin was unaffected by the methanol and heat treatment. Both endogenous liver ferritin and radiolabeled human liver ferritin added to liver homogenates were recovered after methanol and heat treatment with similar yields (77 +/- 7% and 70 +/- 2%, respectively) when compared with the standard treatment of heating a homogenate at 75 degrees C. The overall ferritin yield with this rapid procedure was 40%.
本文描述了一种从人肝组织中纯化铁蛋白的简单、快速技术。在肝匀浆中终浓度为40%（v/v）的甲醇可沉淀大部分蛋白质，但不影响铁蛋白。随后将该匀浆在75℃加热10分钟，通过免疫电泳和聚丙烯酰胺凝胶电泳判断，可得到纯化的铁蛋白制剂。所得纯化铁蛋白所含铁量与原始内源性铁蛋白相同。通过火箭免疫电泳和洛瑞法测量时，纯化铁蛋白制剂中的蛋白量无显著差异（配对t检验），这表明铁蛋白的抗原性不受甲醇和热处理的影响。与在75℃加热匀浆的标准处理相比，甲醇和热处理后，内源性肝铁蛋白和添加到肝匀浆中的放射性标记人肝铁蛋白的回收率相似（分别为77±7%和70±2%）。该快速方法的铁蛋白总产率为40%。
