A key component of efforts to address the reproducibility crisis in biomedical research is the development of rigorously validated and renewable protein-affinity reagents. As part of the US National Institutes of Health (NIH) Protein Capture Reagents Program (PCRP), we have generated a collection of 1,406 highly validated immunoprecipitation- and/or immunoblotting-grade mouse monoclonal antibodies (mAbs) to 737 human transcription factors, using an integrated production and validation pipeline. We used HuProt human protein microarrays as a primary validation tool to identify mAbs with high specificity for their cognate targets. We further validated PCRP mAbs by means of multiple experimental applications, including immunoprecipitation, immunoblotting, chromatin immunoprecipitation followed by sequencing (ChIP-seq), and immunohistochemistry. We also conducted a meta-analysis that identified critical variables that contribute to the generation of high-quality mAbs. All validation data, protocols, and links to PCRP mAb suppliers are available at http://proteincapture.org.

解决生物医学研究中可重复性危机的努力的一个关键组成部分是开发经过严格验证和可再生的蛋白质亲和试剂。作为美国国立卫生研究院 (NIH) 蛋白质捕获试剂计划 (PCRP) 的一部分，我们针对 737 种人类转录因子生成了 1,406 种经过高度验证的免疫沉淀和/或免疫印迹级小鼠单克隆抗体 (mAb)，使用集成的生产和验证管道。我们使用 HuProt 人类蛋白质微阵列作为主要验证工具来鉴定对其同源靶标具有高特异性的 mAb。我们通过多种实验应用进一步验证了 PCRP mAb，包括免疫沉淀、免疫印迹、染色质免疫沉淀后测序 (ChIP-seq) 和免疫组织化学。我们还进行了一项荟萃分析，确定了有助于生成高质量 mAb 的关键变量。所有验证数据、协议和 PCRP mAb 供应商的链接均可在 http://proteincapture.org 上获得。