Department of Neurology and Neurosurgery, Structural Genomics Consortium, The Montreal Neurological Institute, McGill University, Montreal, Québec, H3A 2B4, Canada.
Department of Pharmacology and Toxicology, Indiana University School of Medicine, Indianapolis, Indiana, 46202, USA.
F1000Res. 2024 Jan 18;13:77. doi: 10.12688/f1000research.146156.1. eCollection 2024.
Phosphatidylinositol-specific phospholipase C gamma 2 (PLC-gamma-2) is an enzyme that regulates the function of immune cells. PLC-gamma-2 has been implicated in neurodegenerative and autoimmune disorders, yet investigation of this protein has been limited by a lack of independently characterized antibodies. Here we have characterized eleven PLC-gamma-2 commercial antibodies for use in 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 issues 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.
磷脂酰肌醇特异性磷脂酶 C 伽马 2(PLC-γ-2)是一种调节免疫细胞功能的酶。PLC-γ-2 与神经退行性和自身免疫性疾病有关,但由于缺乏独立特征化的抗体,对该蛋白的研究受到限制。在这里,我们使用标准化的实验方案,通过比较敲除细胞系和同基因亲本对照中的读出值,对 11 种 PLC-γ-2 商业抗体进行了用于 Western Blot、免疫沉淀和免疫荧光的特性分析。这些研究是一个更大的合作计划的一部分,该计划旨在通过对人类蛋白质的商业可用抗体进行特征分析,并将结果公开作为科学界的资源来解决抗体重现性问题。虽然抗体和方案在不同实验室之间有所不同,但我们鼓励读者将本报告用作指南,根据自己的具体需求选择最合适的抗体。