Ayoubi Riham, Fotouhi Maryam, Alende Charles, Southern Kathleen, Laflamme Carl
Department of Neurology and Neurosurgery, Structural Genomics Consortium, The Montreal Neurological Institute, McGill University, Montreal, Québec, H3A 2B4, Canada.
F1000Res. 2024 Jul 9;13:1. doi: 10.12688/f1000research.145146.2. eCollection 2024.
Protein phosphatase 2A is a serine/threonine phosphatase with activity dependent on an associated regulatory subunit, serine/threonine-protein phosphatase 2A 56 kDa regulatory subunit delta (δ) isoform (PPP2R5D). PPP2R5D is the δ isoform in the B56 family of regulatory subunits. Abundantly expressed in the brain and involved in a broad range of cellular processes, PPP2R5D plays an essential role in modulating key neuronal pathways and signalling. Pathogenic mutations in the gene are linked to clinical symptoms characterized by neurodevelopmental delay, intellectual disability, and autism spectrum disorders. The etiology of these genetic disorders remains unknown, which can partly be due to the lack of independently characterized antibodies. Here we have characterized six PPP2R5D commercial antibodies for Western Blot, immunoprecipitation, and immunofluorescence using a standardized experimental protocol based on comparing read-outs in knockout cell lines and isogenic parental controls. These studies are part of a larger, collaborative initiative seeking to address antibody reproducibility by characterizing commercially available antibodies for human proteins and publishing the results openly as a resource for the scientific community. While use of antibodies and protocols vary between laboratories, we encourage readers to use this report as a guide to select the most appropriate antibodies for their specific needs.
蛋白磷酸酶2A是一种丝氨酸/苏氨酸磷酸酶,其活性依赖于相关的调节亚基,即丝氨酸/苏氨酸蛋白磷酸酶2A 56 kDa调节亚基δ(δ)同工型(PPP2R5D)。PPP2R5D是调节亚基B56家族中的δ同工型。PPP2R5D在大脑中大量表达,参与广泛的细胞过程,在调节关键神经元通路和信号传导中起重要作用。该基因的致病突变与以神经发育迟缓、智力残疾和自闭症谱系障碍为特征的临床症状相关。这些遗传疾病的病因仍然未知,部分原因可能是缺乏独立表征的抗体。在这里,我们使用基于比较敲除细胞系和同基因亲本对照读数的标准化实验方案,对六种用于蛋白质印迹、免疫沉淀和免疫荧光的PPP2R5D商业抗体进行了表征。这些研究是一项更大的合作计划的一部分,该计划旨在通过表征人源蛋白质的商业可用抗体并将结果作为科学界的资源公开出版来解决抗体的可重复性问题。虽然不同实验室使用的抗体和方案各不相同,但我们鼓励读者将本报告作为指南,为其特定需求选择最合适的抗体。