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鉴定用于 Western blot、免疫沉淀和免疫荧光的 Sequestosome-1 高活性抗体。

The identification of high-performing antibodies for Sequestosome-1 for use in Western blot, immunoprecipitation and immunofluorescence.

机构信息

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.

出版信息

F1000Res. 2024 Jul 10;12:324. doi: 10.12688/f1000research.132628.2. eCollection 2023.

Abstract

Sequestosome-1, encoded by the gene , functions as a bridge between ubiquitinated proteins and the proteasome or autophagosome, thereby regulating protein degradation pathways. Loss of Sequestosome-1 is hypothesized to enhance neurodegeneration progression in several diseases, including amyotrophic lateral sclerosis (ALS) and frontotemporal disorders (FTD). Sequestosome-1 reproducible research would be facilitated with the availability of well-characterized anti-Sequestosome-1 antibodies. In this study, we characterized seventeen Sequestosome-1 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. 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.

摘要

自噬体相关蛋白 1(Sequestosome-1),由 基因编码,作为连接泛素化蛋白和蛋白酶体或自噬体的桥梁,从而调节蛋白降解途径。自噬体相关蛋白 1 的缺失被假设会增强几种疾病(包括肌萎缩性侧索硬化症(ALS)和额颞叶痴呆(FTD))的神经退行性进程。如果有特征明确的抗自噬体相关蛋白 1 抗体,那么自噬体相关蛋白 1 的可重复性研究将会得到促进。在这项研究中,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的读出值,对 17 种商业抗自噬体相关蛋白 1 抗体进行了 Western blot、免疫沉淀和免疫荧光分析。我们确定了许多性能良好的抗体,并鼓励读者将本报告作为选择最适合其特定需求的抗体的指南。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/23c2/11240125/e61c0950fc26/f1000research-12-168891-g0000.jpg

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