A proteomic approach to cellulose-synthesizing membrane protein complexes in the ascidian Ciona intestinalis

我们报告了一种western blotting系统来评估海鞘中纤维素合成膜蛋白复合物。制备兔多克隆抗体，针对肠梭菌纤维素合成酶Ci-CesA部分合成多肽。在用亲和柱纯化多肽和用Ciona丙酮粉吸附后，非特异性反应性或交叉反应性降低，因此仅在微粒体制备中获得具有估计分子质量的Ci-CesA单条带。在样品准备方面，法压机对乔娜胚胎很有效。

We report a western blotting system to assess cellulose-synthesizing membrane protein complexes in the ascidian Ciona intestinalis. Rabbit polyclonal antibodies were raised against several synthetic peptides corresponding to the parts of Ci-CesA, which is the cellulose synthase of C. intestinalis. After purifications with affinity columns using the peptides and an adsorption with Ciona acetone powder, either non-specific reactivity or cross-reactivity was reduced so that the single band was obtained with the estimated molecular mass of Ci-CesA, only in microsome preparations. For sample preparations, French-press worked fine for Ciona embryos.