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本文研究利用高分子量谷蛋白亚基1Dy10特异性单克隆抗体测定小麦胚乳贮藏蛋白中是否含有优质亚基1Dx5+1Dy10的免疫化学测定条件,建立适宜的间接ELISA测定法。结果表明:蛋白质提取剂种类对籽粒贮藏蛋白提取效率及1Dx5+1Dy10与1Dx2+1Dy12亚基的识别起决定作用。其中以10mMHCl较理想,还原剂DTT能提高1Dx5+1Dy10与1Dx2+1Dy12亚基的识别能力,蛋白提取时间、醇溶蛋白含量以及封闭液的种类都对实验结果具有重要影响。
In this study, the use of high molecular weight glutenin subunit 1Dy10 specific monoclonal antibody to determine whether the wheat endosperm storage protein contains high-quality subunits 1Dx5 + 1Dy10 immunochemical assay conditions, the establishment of appropriate indirect ELISA assay. The results showed that the type of protein extractant played an important role in the extraction efficiency of grain storage protein and the identification of 1Dx5 + 1Dy10 and 1Dx2 + 1Dy12 subunits. Among them, 10mM HCl was the best one. The reducing agent DTT could improve the recognition ability of 1Dx5 + 1Dy10 and 1Dx2 + 1Dy12 subunits, the time of protein extraction, the content of gliadin and the type of blocking solution all had important influence on the experimental results.