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Rapid isolation and identification of individual microvillous proteins from a nitrocellulose membrane filter


, : Rapid isolation and identification of individual microvillous proteins from a nitrocellulose membrane filter. Membrane 19(5): 339-343

Several organic solvents were used to dissolve. the nitrocellulose filter to- which proteins were bound. Among these solvents, acetone and methanol easily dissolved the nitrocellulose. Proteins released from the nitrocellulose were precipitated by centrifugation. The efficiency of protein recovery was investigated using commercially available proteins. In this study, we found that a 50% acetone-50% methanol was most effective to recover the intestinal microvillous membrane proteins from a nitrocellulose filter. Subsequent digestion of these proteins with CNBr, or some proteinases, exposed internal fragments and enabled their microsequencing.

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