Establishment of A Two Dimensional Electrophoresis Protocol for Protein of Gentiana manshurica Flower Bud
An extraction method of total protein from Gentiana manshurica Kitag. flower bud and the technology of two-dimensional electrophoresis of flower bud proteome were initially established in order to raise resolution and repeatability. The preparation and dissolution of samples were optimized, which solved the interference of pigment, polysaccharide, and phenol materials to the two-dimensional electrophoresis disturbance in the process of extraction of G. manshurica flower bud. A kind of effective protein dissolution fluid was determined as 7 mol/L urea, 2 mol/L thiourea, 4% CHAPS, 40 mmol/L DTT, and 2% ampholyte pH 3~10. The optimization of other links in electrophoresis was also conducted. After the Coo-massie Brilliant Blue staining, it could distinguish approximately 1 000 protein spots. A two-dimensional electrophoresis atlas with good repeatability and high resolution was obtained.
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NSTL
万方数据
维普
