Quantitation of human plasma proteins separated on non-denaturing 2-D gels by image analysis.
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概要
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The procedure to quantitate human plasma proteins separated by non-denaturing two-dimensional gel electrophoresis (2-DE) was examined using micro size gels and a commercially available software of image analysis. Varying volumes of a human plasma sample were loaded on micro 2-DE gels and the stained gels were subjected to image acquisition and image analysis. Plasma proteins were identified on the 2-DE gels referring to an "identification map, " which has been prepared by blotting-immunochemical staining. Integrated density and loaded volume of plasma were linearly related for most of the spots, but saturation of integrated density was observed for albumin and some major plasma proteins. Various quantities (0.5-100μg) of purified human albumin were subjected to micro 2-DE and image analysis, and a quantity standard curve was prepared. The quantities and relative contents of plasma proteins, calculated from the standard curve as albumin equivalent values, generally coincided with those reported in literatures.
- 日本電気泳動学会の論文
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