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用于蛋白质免疫印迹、免疫沉淀和免疫荧光的VAPB(通用蛋白质数据库编号:O95292)高性能抗体选择指南。

A guide to selecting high-performing antibodies for VAPB (UniProt ID: O95292) for use in western blot, immunoprecipitation, and immunofluorescence.

作者信息

González Bolívar Sara, Ayoubi Riham, Alende Charles, Fothouhi Maryam, Shlaifer Irina, McPherson Peter S, Laflamme Carl

机构信息

Montreal Neurological Institute-Hospital, Montreal, Québec, Canada.

出版信息

F1000Res. 2024 Dec 24;13:1559. doi: 10.12688/f1000research.160226.1. eCollection 2024.

Abstract

VAPB is an adaptor protein known for its role as an anchor for other proteins at the endoplasmic reticulum. A mutant form of VAPB has been linked to amyotrophic lateral sclerosis and the underlying mechanisms resulting from this defect are studied by researchers in this area to uncover its implication in the disease. Here we have characterized six VAPB 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

VAPB是一种衔接蛋白,因其在内质网中作为其他蛋白的锚定物而闻名。VAPB的一种突变形式与肌萎缩侧索硬化症有关,该领域的研究人员对这种缺陷产生的潜在机制进行了研究,以揭示其在该疾病中的作用。在此,我们使用标准化实验方案,通过比较基因敲除细胞系和同基因亲本对照中的检测结果,对六种用于蛋白质免疫印迹、免疫沉淀和免疫荧光的商业化VAPB抗体进行了表征。这些研究是一个更大的合作项目的一部分,该项目旨在通过对商业化的人类蛋白抗体进行表征并将结果公开作为科学界的资源,来解决抗体可重复性问题。虽然不同实验室使用的抗体和实验方案各不相同,但我们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。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a30f/11799760/342b28164f54/f1000research-13-176085-g0000.jpg

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