Department of Neurology and Neurosurgery, Structural Genomics Consortium, The Montreal Neurological Institute, McGill University, Montreal, Québec, H3A 2B4, Canada.
The Neuro's Early Drug Discovery Unit (EDDU), Structural Genomics Consortium, McGill University, Montreal, Québec, H3A 2B4, Canada.
F1000Res. 2023 Jul 25;12:884. doi: 10.12688/f1000research.139755.1. eCollection 2023.
Charged multivesicular body protein 2B is a subunit of the endosomal sorting complex required for transport III (ESRCT-III), a complex implicated in the lysosomal degradation pathway and formation of multivesicular bodies. Mutations to the gene can result in abnormal protein aggregates in neurons and is therefore predicted to be associated in neurodegenerative diseases, including across the ALS-FTD spectrum. Through our standardized experimental protocol which compares read-outs in knockout cell lines and isogenic parental controls, this study aims to enhance the reproducibility of research on this target by characterizing eight commercial antibodies against charged multivesicular body protein 2b using Western Blot, immunoprecipitation, and immunofluorescence. We identified many high-performing antibodies and encourage readers to use this report as a guide to select the most appropriate antibody for their specific needs.
带电荷的多泡体蛋白 2B 是内体分选复合物必需的亚基 III（ESRCT-III），该复合物参与溶酶体降解途径和多泡体的形成。基因的突变可导致神经元中异常的蛋白质聚集，因此预计与神经退行性疾病有关，包括 ALS-FTD 谱系。通过我们的标准化实验方案，该方案比较了敲除细胞系和同基因亲本对照的读出值，本研究旨在通过使用 Western Blot、免疫沉淀和免疫荧光技术对针对带电荷的多泡体蛋白 2B 的八种商业抗体进行表征，从而提高针对该靶标的研究的可重复性。我们确定了许多性能优异的抗体，并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。
