1. A toolbox of immunoprecipitation-grade monoclonal antibodies to human transcription factors
- Author
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Leonardo Ramos, Kimberly Ruiz, Anand Venkataraman, Keven Murphy, Jessica E. McDade, Atul Tandon, Lizhi Jiang, Christian Rosa, Edisa Albino, Luvir Lugo, Mark Mackiewicz, Sarah Keegan, Shaohui Hu, Kun Yang, Gordon Whiteley, Devlina Ghosh, Jimmy de Melo, Daniel Eichinger, Stephen Anderson, Ivan Vargas, Richard G. Saul, Diane Bayron Kain, Zully Ann Rivera-Pacheco, Lin Xue, Guang Song, Gloriner Morell, Gaetano T. Montelione, Pedro Ramos, Brittany Jones, Shuang Liu, Zheng Kuang, Florencia Pauli-Behn, Richard M. Myers, Wendy Y. Yap, Seth Blackshaw, Simona Colantonio, Seva G. Khambadkone, Lillyann Asencio, Luis Nazario, Heng Zhu, David Fenyö, Ignacio Pino, Joel S. Bader, Milanka Stevanovic, Jose Irizarry, Sooyeon Yoo, Jef D. Boeke, Elliot Campbell, Ruth Almodovar, Yana Li, Hongyan Zhang, Javier Rivera, Paolo Mita, Brian S. Clark, and Moises Vargas
- Subjects
0301 basic medicine ,Databases, Factual ,medicine.drug_class ,Immunoprecipitation ,Protein Array Analysis ,Computational biology ,Biology ,Monoclonal antibody ,Biochemistry ,Article ,03 medical and health sciences ,Mice ,medicine ,Animals ,Humans ,Cloning, Molecular ,Molecular Biology ,Transcription factor ,Mice, Inbred BALB C ,Extramural ,Antibodies, Monoclonal ,Reproducibility of Results ,Cell Biology ,030104 developmental biology ,Protein microarray ,biology.protein ,Female ,Antibody ,Chromatin immunoprecipitation ,Biotechnology ,HeLa Cells ,Transcription Factors - Abstract
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.
- Published
- 2017