Well-characterized antibody reagents play a key role in the reproducibility of research findings, and inconsistent antibody performance leads to variability in Western blotting and other immunoassays. The current lack of clear, accepted standards for antibody validation and reporting of experimental details contributes to this problem. Because the performance of primary antibodies is strongly influenced by assay context, recommendations for validation and usage are unique to each type of immunoassay. Practical strategies are proposed for the validation of primary antibody specificity, selectivity, and reproducibility using Western blot analysis. The antibody should produce reproducible results within and between Western blotting experiments and the observed effect confirmed with a complementary or orthogonal method. Routine implementation of standardized antibody validation and reporting in immunoassays such as Western blotting may promote improved reproducibility across the global life sciences community.

中文翻译：

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免疫印迹的抗体验证：由用户决定。

表征良好的抗体试剂在研究结果的可重复性中起关键作用，并且不一致的抗体性能会导致蛋白质印迹和其他免疫分析的可变性。当前缺乏明确的，可接受的抗体验证标准和实验细节报告的标准导致了这一问题。由于一抗的性能受测定环境的强烈影响，因此对于每种免疫测定类型，验证和使用的建议都是独特的。提出了使用Western blot分析验证一抗特异性，选择性和重现性的实用策略。该抗体应在Western印迹实验之内和之间产生可重复的结果，并通过互补或正交方法确认观察到的效果。