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用于 Western Blot、免疫沉淀和免疫荧光的淀粉样β前体蛋白的高表现抗体选择指南。

A guide to selecting high-performing antibodies for amyloid-beta precursor protein 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.

出版信息

F1000Res. 2024 Apr 10;12:956. doi: 10.12688/f1000research.139867.2. eCollection 2023.

Abstract

The amyloid-beta precursor protein is a transmembrane protein expressed in many tissues and highly concentrated in the brain. The protein is of significant interest due to its involvement in the generation of amyloidogenic β-amyloid peptides, prone to plaque formation that is characteristic of Alzheimer's Disease. The scientific community would benefit from the availability of high-quality anti-amyloid-beta precursor protein antibodies to enhance reproducible research on this target. In this study, we characterized eleven amyloid-beta precursor protein 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.

摘要

淀粉样β前体蛋白是一种跨膜蛋白,在许多组织中表达,在大脑中高度集中。由于其参与生成淀粉样β肽,易形成阿尔茨海默病特征性的斑块,该蛋白引起了广泛关注。科学界将受益于高质量的抗淀粉样β前体蛋白抗体的可用性,以增强对该靶点的可重复研究。在这项研究中,我们使用基于比较敲除细胞系和同基因亲本对照的读出值的标准化实验方案,对 11 种淀粉样β前体蛋白商业抗体进行了 Western blot、免疫沉淀和免疫荧光分析。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抗体进行特征描述,并将结果公开作为科学界的资源,来解决抗体重现性问题。虽然不同实验室之间的抗体和方案使用情况有所不同,但我们鼓励读者将本报告用作指南,根据其特定需求选择最合适的抗体。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c2b5/11445668/52d865ec261a/f1000research-12-164529-g0000.jpg

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