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用于 Western blot、免疫沉淀和免疫荧光实验的 Huntingtin(UniProt ID:P42858)抗体选择指南

A guide to selecting high-performing antibodies for Huntingtin (UniProt ID: P42858) for use in western blot, immunoprecipitation, and immunofluorescence.

机构信息

Structural Genomics Consortium, University of Toronto, Toronto, Ontario, Canada.

Department of Neurology and Neurosurgery, Structural Genomics Consortium, The Montreal Neurological Institute, McGill University, Montreal, Québec, Canada.

出版信息

F1000Res. 2025 Jan 2;13:922. doi: 10.12688/f1000research.153670.1. eCollection 2024.

Abstract

Huntingtin encodes a 3144 amino acid protein, with a polyglutamine repeat tract at the N-terminus. Expansion of this repeat tract above a pathogenic threshold of 36 repeats is the causative mutation of Huntington's disease, a neurodegenerative disorder characterized by loss of striatal neurons. Here we have characterized twenty Huntingtin 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

亨廷顿蛋白编码一个 3144 个氨基酸的蛋白质,其 N 端有一个多聚谷氨酰胺重复序列。这个重复序列的扩展超过 36 个重复的致病阈值是亨廷顿病的致病突变,亨廷顿病是一种神经退行性疾病,其特征是纹状体神经元的丧失。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读出值的标准化实验方案,对二十种亨廷顿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使用的抗体和方案有所不同,但我们鼓励读者将本报告用作指南,根据自己的具体需求选择最合适的抗体。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2476/11801663/6602956a3706/f1000research-13-176211-g0000.jpg

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