| Literature DB >> 29638227 |
Anand Venkataraman1, Kun Yang2, Jose Irizarry3, Mark Mackiewicz4, Paolo Mita5,6,7, Zheng Kuang6, Lin Xue1, Devlina Ghosh1, Shuang Liu8, Pedro Ramos3, Shaohui Hu3, Diane Bayron Kain3, Sarah Keegan5,6, Richard Saul8, Simona Colantonio9, Hongyan Zhang8, Florencia Pauli Behn4, Guang Song8, Edisa Albino3, Lillyann Asencio3, Leonardo Ramos3, Luvir Lugo3, Gloriner Morell3, Javier Rivera3, Kimberly Ruiz3, Ruth Almodovar3, Luis Nazario3, Keven Murphy3, Ivan Vargas3, Zully Ann Rivera-Pacheco3, Christian Rosa3, Moises Vargas3, Jessica McDade7, Brian S Clark1, Sooyeon Yoo1, Seva G Khambadkone10, Jimmy de Melo1, Milanka Stevanovic1, Lizhi Jiang1, Yana Li11, Wendy Y Yap3, Brittany Jones12, Atul Tandon12, Elliot Campbell13,14, Gaetano T Montelione13,14, Stephen Anderson13,14, Richard M Myers4, Jef D Boeke5,6,7, David Fenyö5,6, Gordon Whiteley9, Joel S Bader2, Ignacio Pino3, Daniel J Eichinger3, Heng Zhu8,15, Seth Blackshaw1,15,16,17,18.
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.Entities:
Mesh：
Substances：
Year: 2018 PMID： 29638227 PMCID： PMC6063793 DOI： 10.1038/nmeth.4632
Source DB: PubMed Journal: Nat Methods ISSN： 1548-7091 Impact factor: 28.547