PDF(Pubmed)
Abstract：
Many proteins self-assemble into dimers and higher-order oligomers. Therefore, the goal of this protocol is to characterize the conformational states of an endogenous protein of interest. Here, we present a protocol for assessing protein self-assembly in cell lysates using chemical cross-linking. We describe steps for chemical cross-linking with recombinant proteins as well as steps for cell culture and cell lysate preparation, chemical cross-linking, SDS-PAGE, and western blotting for the detection of endogenous proteins. For complete details on the use and execution of this protocol, please refer to Balaji et al.1.
摘要：
许多蛋白质自组装成二聚体和高阶寡聚体。因此,该方案的目标是表征感兴趣的内源性蛋白质的构象状态.这里,我们提出了一种使用化学交联评估细胞裂解物中蛋白质自组装的方案。我们描述了用重组蛋白进行化学交联的步骤，以及细胞培养和细胞裂解物制备的步骤。化学交联,SDS-PAGE,和蛋白质印迹检测内源性蛋白。有关此协议的使用和执行的完整详细信息，请参考Balaji等人1。
