Zhang Jian-Guo, Nicholson Sandra E
Cancer and Haematology Division, Walter and Eliza Hall Institute of Medical Research, Parkville, VIC, Australia.
Methods Mol Biol. 2013;967:249-59. doi: 10.1007/978-1-62703-242-1_18.
The suppressors of cytokine signalling (SOCS) protein family consist of eight members (SOCS 1-7, and CIS). SOCS1 and SOCS3 are the best-studied family members and have been shown to act as negative feedback inhibitors of the JAK/STAT signalling pathway. To study the physiological roles of the SOCS proteins, it is necessary to establish methods for detecting endogenous proteins often expressed at low levels in cells after cytokine induction. To facilitate the detection of endogenous SOCS1 and SOCS3 proteins, we have generated in-house antibodies specific to these proteins, which we have used together with commercially available antibodies. Here, we describe the methods for immunoprecipitating SOCS1 and SOCS3 proteins from mouse tissue extracts and their subsequent detection by Western blot analysis. These methods can also be applied to the detection of SOCS1 and SOCS3 in cell lines.
细胞因子信号转导抑制因子（SOCS）蛋白家族由八个成员组成（SOCS 1 - 7和CIS）。SOCS1和SOCS3是研究最为深入的家族成员，已被证明可作为JAK/STAT信号通路的负反馈抑制剂。为了研究SOCS蛋白的生理作用，有必要建立检测细胞因子诱导后细胞中常以低水平表达的内源性蛋白的方法。为便于检测内源性SOCS1和SOCS3蛋白，我们制备了针对这些蛋白的内部抗体，并将其与市售抗体一起使用。在此，我们描述从小鼠组织提取物中免疫沉淀SOCS1和SOCS3蛋白的方法，以及随后通过蛋白质印迹分析进行检测的方法。这些方法也可应用于细胞系中SOCS1和SOCS3的检测。
