RT Journal Article SR Electronic T1 A toolbox of immunoprecipitation-grade monoclonal antibodies against human transcription factors JF bioRxiv FD Cold Spring Harbor Laboratory SP 116442 DO 10.1101/116442 A1 Anand Venkataraman A1 Kun Yang A1 Jose Irizarry A1 Mark Mackiewicz A1 Paolo Mita A1 Zheng Kuang A1 Lin Xue A1 Devlina Ghosh A1 Shuang Liu A1 Pedro Ramos A1 Shaohui Hu A1 Diane Bayron A1 Sarah Keegan A1 Richard Saul A1 Simona Colantonio A1 Hongyan Zhang A1 Florencia Pauli Behn A1 Guang Song A1 Edisa Albino A1 Lillyann Asencio A1 Leonardo Ramos A1 Luvir Lugo A1 Gloriner Morell A1 Javier Rivera A1 Kimberly Ruiz A1 Ruth Almodovar A1 Luis Nazario A1 Keven Murphy A1 Ivan Vargas A1 Zully Ann Rivera-Pacheco A1 Christian Rosa A1 Moises Vargas A1 Jessica McDade A1 Brian S. Clark A1 Sooyeon Yoo A1 Seva G. Khambadkone A1 Jimmy de Melo A1 Milanka Stevanovic A1 Lizhi Jiang A1 Yana Li A1 Wendy Y. Yap A1 Brittany Jones A1 Atul Tandon A1 Elliot Campbell A1 Stephen Anderson A1 Richard M. Myers A1 Jef D. Boeke A1 David Fenyo A1 Gordon Whiteley A1 Joel S. Bader A1 Ignacio Pino A1 Daniel J. Eichinger A1 Heng Zhu A1 Seth Blackshaw YR 2017 UL http://biorxiv.org/content/early/2017/03/16/116442.abstract AB A key component to overcoming the reproducibility crisis in biomedical research is the development of readily available, rigorously validated and renewable protein affinity reagents. As part of the NIH Protein Capture Reagents Program (PCRP), we have generated a collection of 1406 highly validated, immunoprecipitation (IP) and/or immunoblotting (IB) grade, mouse monoclonal antibodies (mAbs) to 736 human transcription factors. We used HuProtâ„¢ human protein microarrays to identify mAbs that recognize their cognate targets with exceptional specificity. Using an integrated production and validation pipeline, we validated these mAbs in multiple experimental applications, and have distributed them to the Developmental Studies Hybridoma Bank (DSHB) and several commercial suppliers. This study allowed us to perform a meta-analysis that identified critical variables that contribute to the generation of high quality mAbs. We find that using full-length antigens for immunization, in combination with HuProtâ„¢ analysis, provides the highest overall success rates. The efficiencies built into this pipeline ensure substantial cost savings compared to current standard practices.