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Extraction and separation of cell supernatant protein
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Cell supernatant protein was extracted by the standard methanol/chloroform method. The cell supernatants were layered with 1/4 volume of chloroform, and the same volume of methanol was added. After evenly mixing the solutions on a vortex shaker, the mixture was centrifuged at 12,000 rpm for 5 min at room temperature. The upper aqueous phase (methanol) was carefully removed, and the intermediate protein layer was aspirated and extracted once with methanol as described above.
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