在TCA/丙酮沉淀法的基础上，用乙醇和乙醚洗涤（称双乙法）沉淀获得橡胶树胶乳C-乳清蛋白样品，并在样品溶解时通过超声助溶进一步提高样品的溶解度。结果显示，利用改良后的样品制备方法（TCA/丙酮+双乙法+超声处理）所获得的C-乳清蛋白样品的溶解性大幅增加，蛋白得率高达8.47 mg/mL C-乳清，为传统TCA/丙酮法的1.6倍；同时降低了盐离子浓度，等电聚焦时升压顺利、聚焦完全，双向电泳的图谱质量明显改善，在银染2D胶上可获得1 206个蛋白点，为传统TCA/丙酮法的2.7倍。本文对胶乳C-乳清蛋白样品制备方法进行深入优化，获得了重现性较好的高质量双向电泳蛋白图谱，促进了橡胶树蛋白质组学研究。
The C-serum is a major component of the latex and a key material for proteinomics of Hevea brasiliensis. Using the 2-D electrophoresis, there are some disadvantages with protein extraction from C-serum, such as unsatisfied solubility, higher ions concentrations and too many impurities. In this paper, the alcohol-ether method and sonication were used to further remove interfering substances and solubilized protein based on the traditional method of TCA/acetone. Comparing the traditional method of extraction, the results showed that the new method improved the solubility, ions concentration and purity of protein samples from C-serum. The optimized methods for extraction of protein will be extremely useful for the proteomic research of C-serum form H. brasiliensis latex.
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