METHOD OF CYTOCHROME C PREPARING Russian patent published in 1997 - IPC

FIELD: biochemistry. SUBSTANCE: invention relates to isolation of cytochrome c from the cattle hearts. Vessels and ligaments were removed, heart tissue is defatted, milled and blood is washed out. The obtained homogenate is extracted with an aqueous solution of 0.10-0.15% sulfuric acid or 2.0-2.5% trichloroacetic acid at 10-15 C, at pH = 3.9-4.2. Extract is alkalinized to pH = 6.8-7.4 and sorption of the end product is carried out on carboxyl-cationite that is a polymer of an order that includes copolymer of methacrylic acid with N,N'-hexamethylenedimethacrylamide and copolymer of acrylic acid with divinyl benzene. Cationite is used in Na⁺-form. Elution is carried out with 10-12% ammonium sulfate solution at pH = 9.0-9.8. Inert proteins were precipitated from eluate by addition of ammonium sulfate at amount 0.5 g/ml elute at pH = 7.2-7.5. Cytochrome c is precipitated from supernatant with 20% trichloroacetic acid solution at cooling. Precipitate is dissolved in small water volume and dialyzed against isotonic solution of sodium chloride up to complete ions SO2-4