Cutts Erin E, Taylor Gillian C, Pardo Mercedes, Yu Lu, Wills Jimi C, Choudhary Jyoti S, Vannini Alessandro, Wood Andrew J
Cancer Biology Division, Institute of Cancer Research, London, SW7 3RP, UK.
MRC Human Genetics Unit, Institute of Genetics and Molecular Medicine, University of Edinburgh, Edinburgh, EH4 2XU, UK.
Wellcome Open Res. 2021 Jan 13;6:3. doi: 10.12688/wellcomeopenres.16482.1. eCollection 2021.
Condensin complexes compact and disentangle chromosomes in preparation for cell division. Commercially available antibodies raised against condensin subunits have been widely used to characterise their cellular interactome. Here we have assessed the specificity of a polyclonal antibody (Bethyl A302-276A) that is commonly used as a probe for NCAPH2, the kleisin subunit of condensin II, in mammalian cells. We find that, in addition to its intended target, this antibody cross-reacts with one or more components of the SWI/SNF family of chromatin remodelling complexes in an NCAPH2-independent manner. This cross-reactivity, with an abundant chromatin-associated factor, is likely to affect the interpretation of protein and chromatin immunoprecipitation experiments that make use of this antibody probe.
凝聚素复合物在细胞分裂准备过程中使染色体浓缩并解开缠结。针对凝聚素亚基产生的商业可用抗体已被广泛用于表征其细胞相互作用组。在这里，我们评估了一种多克隆抗体（Bethyl A302-276A）的特异性，该抗体通常用作哺乳动物细胞中凝聚素II的kleisin亚基NCAPH2的探针。我们发现，除了其预期靶点外，该抗体还以不依赖NCAPH2的方式与染色质重塑复合物的SWI/SNF家族的一种或多种成分发生交叉反应。这种与丰富的染色质相关因子的交叉反应可能会影响利用该抗体探针进行的蛋白质和染色质免疫沉淀实验的解释。
