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A toolbox of immunoprecipitation-grade monoclonal antibodies to human transcription factors

2018; Nature Portfolio; Volume: 15; Issue: 5 Linguagem: Inglês

10.1038/nmeth.4632

1548-7105

Anand Venkataraman, Kun Yang, José Irizarry, Mark Mackiewicz, Paolo Mita, Zheng Kuang, Xue Lin, D. Ghosh, Shuang Liu, Pedro Luis Ramos‐González, Shaohui Hu, Diane Bayron Kain, Sarah Keegan, Richard G. Saul, Simona Colantonio, Hongyan Zhang, Florencia Pauli Behn, Guang Song, Edisa Albino, Lillyann Asencio, Leonardo Ramos, Luvir Lugo, Gloriner Morell, Javier Sandoval, Kimberly Ruiz, Ruth Almodóvar, Luis Nazario, Keven Murphy, Ivan Vargas, Zully Ann Rivera-Pacheco, Christian Rosa, Moises Vargas, Jessica E. McDade, Brian S. Clark, Sooyeon Yoo, Seva G. Khambadkone, Jimmy de Melo, Milanka Stevanovic, Lizhi Jiang, Yana Li, W Y Yap, Brittany R. Jones, Atul K. Tandon, Elliot Campbell, G.T. Montelione, Stephen K. Anderson, R Myers, Jef D. Boeke, David Fenyö, Gordon Whiteley, Joel S. Bader, Ignacio Pino, Daniel Eichinger, Heng Zhu, Seth Blackshaw,

Viral Infectious Diseases and Gene Expression in Insects

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.