Separation of microsomal cytochrome b5 via phase separation in a mixed solution of Triton X-114 and charged dextran

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The successful introduction of a charged dextran into the Triton X-114 phase separation system for the selective extraction of cytochrome b 5 (cyt. b 5) in liver microsomes is described. In the absence of charged dextran, 55% of total microsomal proteins and 84% of cyt. b 5 were extracted into the surfactant-rich phase. In the presence of anionic dextran sulfate, the extractability of total microsomal proteins was greatly reduced while that of cyt. b 5 was increased. After triplicate extraction, cyt. b 5 was purified more than 10-fold from microsomes with a recovery of 91% in the surfactant-rich phase. In view of its operational simplicity, this method provides a good means for the partial purification of cyt. b 5 prior to chromatographic separations.