A procedure for the purification of ferritin from human liver by heating a methanol-treated homogenate.
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%.
D002851:Chromatography, High Pressure Liquid / Q000379:methods; D004587:Electrophoresis, Agar Gel; D004591:Electrophoresis, Polyacrylamide Gel; D005293:Ferritins / Q000302:isolation & purification*; D006358:Hot Temperature; D006801:Humans; D007122:Immunoelectrophoresis; D007457:Iodine Radioisotopes; D008099:Liver / Q000032:analysis; D000432:Methanol
D007457:Iodine Radioisotopes; D005293:Ferritins; D000432:Methanol