Searching for interactors of endogenous SLX4

The project was aimed at identifying binding partners of endogenous SLX4 in HeLa cells. Whole cell extracts from HeLa Flp-In T-Rex cells were used for immunoprecipitation performed with anti-SLX4 antibodies from Bethyl Laboratories (A302-270A and A302-269A), either used alone or in combination. To discriminate bona fide SLX4 partners from non-specific proteins recognized by the anti-SLX4 antibodies, cells were transfected with control (Luc) siRNA or a siRNA targeting SLX4. After immunoprecipitation, protein complexes were analysed by LC-MSMS.

本项目旨在鉴定HeLa细胞中内源性SLX4的结合伴侣蛋白。本研究采用HeLa Flp-In T-Rex细胞的全细胞提取物，使用Bethyl Laboratories公司的抗SLX4抗体（货号：A302-270A、A302-269A）开展免疫沉淀实验，抗体可单独使用或联合应用。为区分SLX4的特异性结合蛋白与抗SLX4抗体非特异性识别的杂蛋白，实验将细胞分为两组，分别转染靶向荧光素酶（Luc）的阴性对照小干扰RNA（small interfering RNA，siRNA），或靶向SLX4的siRNA。免疫沉淀完成后，通过液相色谱-串联质谱（Liquid Chromatography-Tandem Mass Spectrometry，LC-MSMS）对蛋白复合物进行分析。