TY - JOUR T1 - A toolbox of immunoprecipitation-grade monoclonal antibodies against human transcription factors JF - bioRxiv DO - 10.1101/116442 SP - 116442 AU - Anand Venkataraman AU - Kun Yang AU - Jose Irizarry AU - Mark Mackiewicz AU - Paolo Mita AU - Zheng Kuang AU - Lin Xue AU - Devlina Ghosh AU - Shuang Liu AU - Pedro Ramos AU - Shaohui Hu AU - Diane Bayron AU - Sarah Keegan AU - Richard Saul AU - Simona Colantonio AU - Hongyan Zhang AU - Florencia Pauli Behn AU - Guang Song AU - Edisa Albino AU - Lillyann Asencio AU - Leonardo Ramos AU - Luvir Lugo AU - Gloriner Morell AU - Javier Rivera AU - Kimberly Ruiz AU - Ruth Almodovar AU - Luis Nazario AU - Keven Murphy AU - Ivan Vargas AU - Zully Ann Rivera-Pacheco AU - Christian Rosa AU - Moises Vargas AU - Jessica McDade AU - Brian S. Clark AU - Sooyeon Yoo AU - Seva G. Khambadkone AU - Jimmy de Melo AU - Milanka Stevanovic AU - Lizhi Jiang AU - Yana Li AU - Wendy Y. Yap AU - Brittany Jones AU - Atul Tandon AU - Elliot Campbell AU - Stephen Anderson AU - Richard M. Myers AU - Jef D. Boeke AU - David Fenyo AU - Gordon Whiteley AU - Joel S. Bader AU - Ignacio Pino AU - Daniel J. Eichinger AU - Heng Zhu AU - Seth Blackshaw Y1 - 2017/01/01 UR - http://biorxiv.org/content/early/2017/03/15/116442.abstract N2 - 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. ER -