ALS-RAP 建立经敲除验证的 ALS 研究抗体资源库
A validated antibody toolbox for ALS research
研究团队建立 ALS-Reproducible Antibody Platform（ALS-RAP），对靶向 33 个 ALS 相关蛋白的 303 个抗体进行敲除验证，筛选出可用于 western blot、免疫沉淀和免疫荧光的高质量抗体。此前该团队用敲除验证流程发现针对 C9orf72 的常用抗体缺乏特异性，并在更大规模评测中发现高达 61% 的抗体未达到厂商推荐性能。利用这批验证抗体，研究者分析了人诱导多能干细胞来源及原代神经细胞类型中的蛋白水平，发现多种 ALS 相关蛋白在胶质细胞群体中水平更高、细胞分布多样，与免疫参与 ALS 的新证据一致。
来源：eLife · doi.org
A substantial fraction of amyotrophic lateral sclerosis (ALS)-associated proteins remains poorly characterized, in part because of the limited availability of validated research antibodies. We established knockout (KO)-based antibody characterization workflows and demonstrated that widely used antibodies against the major ALS-associated protein C9orf72 lacked specificity (Laflamme et al., 2019). We subsequently scaled this framework to systematically benchmark research antibodies, revealing that up to 61% fail to perform as recommended by manufacturers (Ayoubi et al., 2023b). Here, we extend this approach by establishing the ALS-Reproducible Antibody Platform (ALS-RAP), a comprehensive effort to generate a publicly available dataset of KO-validated antibodies targeting proteins encoded by ALS risk genes. In total, we characterized 303 antibodies against 33 ALS-associated proteins to identify high-quality reagents for use in western blot, immunoprecipitation, and immunofluorescence. Using these antibodies, we profiled protein levels across human induced pluripotent stem cell-derived and primary neurological cell types. These analyses revealed diverse cellular distributions and higher levels of several ALS-associated proteins in glial populations, consistent with emerging evidence for immune contributions to ALS. Together, ALS-RAP provides a validated antibody toolbox and protein expression resource to support the study of ALS-associated proteins.