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鉴定用于蛋白质免疫印迹、免疫沉淀和免疫荧光的液泡蛋白分选相关蛋白35(hVPS35)的高效抗体。

Identification of high-performing antibodies for Vacuolar protein sorting-associated protein 35 (hVPS35) for use in Western Blot, immunoprecipitation and immunofluorescence.

作者信息

Ayoubi Riham, Fotouhi Maryam, Southern Kathleen, McPherson Peter S, Laflamme Carl

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. 2023 Oct 16;12:452. doi: 10.12688/f1000research.133696.2. eCollection 2023.

Abstract

Vacuolar protein sorting-associated protein 35 is a subunit of the retromer complex, a vital constituent of the endosomal protein sorting pathway. The D620N mutation in the gene has been reported to be linked to type 17 Parkinson's Disease progression, the exact molecular mechanism remains to be solved. The scientific community would benefit from the accessibility of validated and high-quality anti-hVPS35 antibodies. In this study, we characterized thirteen hVPS35 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

液泡蛋白分选相关蛋白35是逆转录复合物的一个亚基,是内体蛋白分选途径的重要组成部分。据报道,该基因中的D620N突变与17型帕金森病的进展有关,确切的分子机制仍有待解决。科学界将受益于经过验证的高质量抗人VPS35抗体。在本研究中,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13种用于蛋白质免疫印迹、免疫沉淀和免疫荧光的商业化抗人VPS35抗体进行了表征。我们鉴定出了许多高性能抗体,并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/11cf/10905018/a4a7870615e9/f1000research-12-157173-g0000.jpg

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