Department of Pathology, Yale University School of Medicine, New Haven, CT 06520, USA.
Biotechniques. 2010 Mar;48(3):197-209. doi: 10.2144/000113382.
Antibodies are among the most frequently used tools in basic science research and in clinical assays, but there are no universally accepted guidelines or standardized methods for determining the validity of these reagents. Furthermore, for commercially available antibodies, it is clear that what is on the label does not necessarily correspond to what is in the tube. To validate an antibody, it must be shown to be specific, selective, and reproducible in the context for which it is to be used. In this review, we highlight the common pitfalls when working with antibodies, common practices for validating antibodies, and levels of commercial antibody validation for seven vendors. Finally, we share our algorithm for antibody validation for immunohistochemistry and quantitative immunofluorescence.
抗体是基础科学研究和临床检测中最常使用的工具之一，但目前还没有普遍接受的用于确定这些试剂有效性的指南或标准化方法。此外，对于市售抗体，显然标签上的内容不一定与管内的内容相对应。要验证抗体的有效性，必须证明其在预期使用的情况下具有特异性、选择性和可重复性。在这篇综述中，我们强调了使用抗体时常见的陷阱、验证抗体的常见做法以及七个供应商的商业抗体验证水平。最后，我们分享了用于免疫组织化学和定量免疫荧光的抗体验证算法。
