Optimized Mouse-on-mouse Immunohistochemical Detection of Mouse ESR2 Proteins with PPZ0506 Monoclonal Antibody.

IF 16.4 1区化学 Q1 CHEMISTRY, MULTIDISCIPLINARY Accounts of Chemical Research Pub Date : 2022-10-28 Epub Date: 2022-10-25 DOI:10.1267/ahc.22-00043

Mina Ozawa, Yujiro Hattori, Shimpei Higo, Mai Otsuka, Keisuke Matsumoto, Hitoshi Ozawa, Hirotaka Ishii

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引用次数: 4

Abstract

Despite the physiological significance of ESR2, a lack of well-validated detection systems for ESR2 proteins has hindered progress in ESR2 research. Thus, recent identification of a specific anti-human ESR2 monoclonal antibody (PPZ0506) and its specific cross-reactivity against mouse and rat ESR2 proteins heightened momenta toward development of appropriate immunohistochemical detection systems for rodent ESR2 proteins. Building upon our previous optimization of ESR2 immunohistochemical detection in rats using PPZ0506, in this study, we further aimed to optimize mouse-on-mouse immunohistochemical detection using PPZ0506. Our assessment of several staining conditions using paraffin-embedded ovary sections revealed that intense heat-induced antigen retrieval, appropriate blocking, and appropriate antibody dilutions were necessary for optimization of mouse-on-mouse immunohistochemistry. Subsequently, we applied the optimized immunostaining method to determine expression profiles of mouse ESR2 proteins in peripheral tissues and brain subregions. Our analyses revealed more localized distribution of mouse ESR2 proteins than previously assumed. Moreover, comparison of these results with those obtained in humans and rats using PPZ0506 revealed interspecies differences in ESR2 expression. We expect that our optimized methodology for immunohistochemical staining of mouse ESR2 proteins will help researchers to solve multiple lines of controversial evidence concerning ESR2 expression.

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PPZ0506单克隆抗体优化小鼠ESR2蛋白的小鼠对小鼠免疫组化检测

尽管ESR2具有重要的生理意义，但缺乏经过验证的ESR2蛋白检测系统阻碍了ESR2研究的进展。因此，最近发现的一种特异性抗人ESR2单克隆抗体(PPZ0506)及其对小鼠和大鼠ESR2蛋白的特异性交叉反应性，促进了开发适合啮齿动物ESR2蛋白的免疫组织化学检测系统的进程。在我们之前使用PPZ0506优化大鼠ESR2免疫组化检测的基础上，在本研究中，我们进一步旨在优化使用PPZ0506的小鼠对小鼠免疫组化检测。我们对石蜡包埋卵巢切片的几种染色条件进行了评估，结果显示，强烈的热诱导抗原回收、适当的阻断和适当的抗体稀释是优化小鼠对小鼠免疫组织化学的必要条件。随后，我们应用优化后的免疫染色方法测定小鼠ESR2蛋白在外周组织和脑亚区表达谱。我们的分析显示小鼠ESR2蛋白的局部分布比之前假设的要多。此外，将这些结果与使用PPZ0506在人和大鼠中获得的结果进行比较，发现ESR2的表达在种间存在差异。我们期望我们优化的小鼠ESR2蛋白免疫组织化学染色方法将帮助研究人员解决有关ESR2表达的多条有争议的证据。

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来源期刊

Accounts of Chemical Research 化学-化学综合

CiteScore

31.40

自引率

1.10%

发文量

312

审稿时长

2 months

期刊介绍： Accounts of Chemical Research presents short, concise and critical articles offering easy-to-read overviews of basic research and applications in all areas of chemistry and biochemistry. These short reviews focus on research from the author’s own laboratory and are designed to teach the reader about a research project. In addition, Accounts of Chemical Research publishes commentaries that give an informed opinion on a current research problem. Special Issues online are devoted to a single topic of unusual activity and significance. Accounts of Chemical Research replaces the traditional article abstract with an article "Conspectus." These entries synopsize the research affording the reader a closer look at the content and significance of an article. Through this provision of a more detailed description of the article contents, the Conspectus enhances the article's discoverability by search engines and the exposure for the research.

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