Ruíz Moleón Vera, Alende Charles, Fotouhi Maryam, Southern Kathleen, Ayoubi Riham, Laflamme Carl
Department of Neurology and Neurosurgery, Structural Genomics Consortium, The Montreal Neurological Institute, McGill University, Montreal, Québec, H3A 2B4, Canada.
F1000Res. 2024 Sep 17;13:1061. doi: 10.12688/f1000research.156209.1. eCollection 2024.
Rab10 is a small GTPase involved in cargo transport from the trans-Golgi network to the plasma membrane and endocytic recycling back to the cell membrane. It has garnered significant interest in neurodegenerative disease research, particularly due to its phosphorylation by the LRRK2 kinase. This relationship underscores the importance of Rab10 in cellular processes related to disease pathology, specifically Parkinson's disease. The accessibility of renewable and selective antibodies against Rab10 would advance research efforts, enabling further understanding of its function and implications in disease. Here, we have characterized eight Rab10 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 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.
Rab10是一种小GTP酶，参与从反式高尔基体网络到质膜的货物运输以及内吞再循环回到细胞膜的过程。它在神经退行性疾病研究中引起了极大关注，特别是因为它被LRRK2激酶磷酸化。这种关系凸显了Rab10在与疾病病理学相关的细胞过程中的重要性，特别是在帕金森病中。针对Rab10的可再生和选择性抗体的可及性将推动研究工作，有助于进一步了解其在疾病中的功能和影响。在这里，我们使用基于比较敲除细胞系和同基因亲本对照中的读数的标准化实验方案，对八种用于蛋白质印迹、免疫沉淀和免疫荧光的Rab10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针对人类蛋白质的商业可用抗体并将结果公开作为科学界的资源来解决抗体可重复性问题。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。
