ニトロセルロース膜を用いた疎水性クロマトグラフィー

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  • Hydrophobic chromatography of proteins using a nitrocellulose membrane.

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A technique of chromatography using a nitrocellulose (NC) membrane was established for the separation of proteins. The proteins were developed on the NC membrane with an aqueous solution containing a non-ionic detergent, polyoxyethylene sorbitane (Tween 20). The rate of flow of the proteins correlated to the strength of hydrophobicity of the proteins which was theoretically calculated from the hydrophobicity of side chains of amino acid residues composed of the proteins, but neither to the molecular weight nor to the isoelectric point of the proteins. The detailed examinations on the effects of pH, temperature, and chemicals such as urea, neutral salts, methanol, and an ionic detergent (SDS) on the chromatography strongly suggested that hydrophobic interactions between the proteins and nitrocellulose play a major role in the protein binding to nitrocellulose membrane. So that, this technique may be useful for the separation of a very small amount of membrane bound proteins such as enzyme complexes, receptors, and ribosomes into their components.

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