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Author:
Date:
2013-02-05
[Abstract] Co-Immunoprecipitation (Co-IP) is the method used to pull down protein partners of a protein of interest using an antibody that specifically binds to this specific protein in order to test protein-protein interaction. “Pulled down” proteins can be analyzed by western blot for suspected protein partner, or by Mass spectrometry for high throughput protein partner identification. The advantage of this technique is that endogenous protein partners can be identified from cell lines that naturally express these factors.
This protocol is optimized for hard-to-extract nuclear proteins, e.g., that stick to the nuclei inclusion bodies / nucleosome complexes such as TLX1 and TLX3 (Dadi et al., 2012). Most often, these factors are not soluble when using classical protein ...
[摘要] 共免疫沉淀（Co-IP）是用于使用特异性结合该特异性蛋白的抗体来下拉目标蛋白的蛋白质配偶体以测试蛋白质 - 蛋白质相互作用的方法。 "下拉"蛋白质可通过蛋白质印迹分析用于可疑蛋白质伴侣，或通过质谱法用于高通量蛋白质伴侣鉴定。这种技术的优点是可以从天然表达这些因子的细胞系中鉴定内源性蛋白质伴侣。
该方案针对难以提取的核蛋白进行优化，例如到核内含体/核小体复合物例如TLX1和TLX3（Dadi等人，2012）。最常见的是，当使用经典的蛋白质提取方法时，这些因素是不可溶的。我们用来添加核酸酶以增加被包涵体中捕获的蛋白质复合物的溶解;尽管功效根据给定的蛋白质而变化，因此必须根据经验确定。
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