Characterization of protein binding to a nitrocellulose membrane.

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  • タンパク質のニトロセルロース膜への結合特性

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Proteins bound to a nitrocellulose (NC) membrane tightly and quantitatively until the NC membrane was saturated with the proteins. The binding of proteins to the NC membrane was characterized by using twenty proteins having different molecular weight and isoelectric point as follows: 1) hydrophobic interactions between the protein and the NC membrane matrices may play a major role in the protein binding, and sugars, amino acids, DNA, nucleotides, neutral salts, or glycerol in the protein solution did not interfere with the protein binding. 2) The number of the protein molecule bound to the NC membrane was in the range from 1.13 to 1.98nmol/cm2 except for a few small and strongly charged proteins. 3) Non-ionic detergents such as Tween 20, Triton X-100, and Nonidet P-40 strongly interfered with the protein binding to the NC membrane in a concentration dependent manner. 4) Proteins could be extracted from the NC membrane with high yield by using a low concentration of the non-ionic detergent. These enables us to assay the proteins with high sensitivity and reproducibility. Furthermore, it may be possible to determine the amino acid composition and sequence with small amount of the proteins after the assay.

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