A guide to selecting high-performing antibodies for Protein-glutamine gamma-glutamyltransferase 2 (TGM2) for use in western blot, immunoprecipitation and immunofluorescence

Riham Ayoubi, Maryam Fotouhi, Charles Alende, Sara González Bolívar, K. Southern, C. Laflamme
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Abstract

Protein-glutamine gamma-glutamyltransferase 2 (TGM2) is a Ca2+ dependent enzyme that catalyzes transglutaminase cross-linking modifications. TGM2 is involved in various diseases, either in a protective or contributory manner, making it a crucial protein to study and determine its therapeutic potential. Identifying high-performing TGM2 antibodies would facilitate these investigations. Here we have characterized seventeen TGM2 commercial antibodies for western blot and sixteen for immunoprecipitation, and immunofluorescence. The implemented standardized experimental protocol is based on comparing read-outs in knockout cell lines against their isogenic parental controls. This study is 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 the 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.
用于 Western 印迹、免疫沉淀和免疫荧光的蛋白-谷氨酰胺γ-谷氨酰转移酶 2 (TGM2) 高效抗体选择指南
蛋白-谷氨酰胺γ-谷氨酰转移酶 2(TGM2)是一种依赖于 Ca2+ 的酶,可催化转谷氨酰胺酶交联修饰。TGM2 以保护性或促成性方式参与各种疾病,因此是研究和确定其治疗潜力的关键蛋白。找出高效的 TGM2 抗体将有助于这些研究。在此,我们对 17 种 TGM2 商品抗体进行了表征,其中 17 种用于 Western 印迹,16 种用于免疫沉淀和免疫荧光。实施标准化实验方案的基础是比较基因敲除细胞系与同源亲代对照的读出结果。这项研究是一项大型合作计划的一部分,该计划旨在通过鉴定市售的人类蛋白质抗体来解决抗体的可重复性问题,并将结果公开发表,作为科学界的资源。虽然不同实验室使用的抗体和方案各不相同,但我们鼓励读者以本报告为指南,根据自己的具体需要选择最合适的抗体。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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