Department of Physiology, Tulane University School of Medicine, New Orleans, Louisiana, United States.
Department of Physiology, East Carolina University, Greenville, North Carolina, United States.
Am J Physiol Renal Physiol. 2024 Mar 1;326(3):F511-F533. doi: 10.1152/ajprenal.00347.2023. Epub 2024 Jan 18.
Antibodies are one of the most used reagents in scientific laboratories and are critical components for a multitude of experiments in physiology research. Over the past decade, concerns about many biological methods, including those that use antibodies, have arisen as several laboratories were unable to reproduce the scientific data obtained in other laboratories. The lack of reproducibility could be largely attributed to inadequate reporting of detailed methods, no or limited verification by authors, and the production and use of unvalidated antibodies. The goal of this guideline article is to review best practices concerning commonly used techniques involving antibodies, including immunoblotting, immunohistochemistry, and flow cytometry. Awareness and integration of best practices will increase the rigor and reproducibility of these techniques and elevate the quality of physiology research.
抗体是科学实验室中使用最广泛的试剂之一,也是生理学研究中多种实验的关键组成部分。在过去的十年中,由于许多实验室无法复制其他实验室获得的科学数据,人们对包括抗体在内的许多生物方法产生了担忧。缺乏可重复性在很大程度上可归因于对详细方法的报告不充分、作者没有或有限的验证,以及未经验证的抗体的生产和使用。本文指南的目的是回顾涉及抗体的常用技术的最佳实践,包括免疫印迹、免疫组织化学和流式细胞术。对最佳实践的认识和整合将提高这些技术的严谨性和可重复性,并提高生理学研究的质量。