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

被引：51

作者：

Venkataraman, Anand ^{[1
]}

Yang, Kun ^{[2
]}

Irizarry, Jose ^{[3
]}

Mackiewicz, Mark ^{[4
]}

Mita, Paolo ^{[5
,6
,7
]}

Kuang, Zheng ^{[6
,19
]}

Xue, Lin ^{[1
]}

Ghosh, Devlina ^{[1
]}

Liu, Shuang ^{[8
]}

Ramos, Pedro ^{[3
]}

Hu, Shaohui ^{[3
]}

Kain, Diane Bayron ^{[3
,20
]}

Keegan, Sarah ^{[5
,6
]}

Saul, Richard ^{[8
]}

Colantonio, Simona ^{[9
]}

Zhang, Hongyan ^{[8
]}

Pauli-Behn, Florencia ^{[4
]}

Song, Guang ^{[8
]}

Albino, Edisa ^{[3
]}

Asencio, Lillyann ^{[3
]}

Ramos, Leonardo ^{[3
]}

Lugo, Luvir ^{[3
]}

Morell, Gloriner ^{[3
]}

Rivera, Javier ^{[3
]}

Ruiz, Kimberly ^{[3
]}

Almodovar, Ruth ^{[3
]}

Nazario, Luis ^{[3
]}

Murphy, Keven ^{[3
]}

Vargas, Ivan ^{[3
]}

Rivera-Pacheco, Zully Ann ^{[3
]}

Rosa, Christian ^{[3
]}

Vargas, Moises ^{[3
]}

McDade, Jessica ^{[7
]}

Clark, Brian S. ^{[1
]}

Yoo, Sooyeon ^{[1
]}

Khambadkone, Seva G. ^{[10
]}

de Melo, Jimmy ^{[1
]}

Stevanovic, Milanka ^{[1
]}

Jiang, Lizhi ^{[1
]}

Li, Yana ^{[11
]}

Yap, Wendy Y. ^{[3
]}

Jones, Brittany ^{[12
]}

Tandon, Atul ^{[12
]}

Campbell, Elliot ^{[13
,14
]}

Montelione, Gaetano T. ^{[13
,14
]}

Anderson, Stephen ^{[13
,14
]}

Myers, Richard M. ^{[4
]}

Boeke, Jef D. ^{[5
,6
,7
]}

Fenyo, David ^{[5
,6
]}

Whiteley, Gordon ^{[9
]}

机构：

[1] Johns Hopkins Univ, Sch Med, Solomon H Snyder Dept Neurosci, Baltimore, MD 21205 USA

[2] Johns Hopkins Univ, Sch Med, Dept Biomed Engn, Baltimore, MD 21205 USA

[3] CDI Labs, Mayaguez, PR 00682 USA

[4] HudsonAlpha Inst Biotechnol, Huntsville, AL USA

[5] NYU Langone Hlth, Inst Syst Genet, New York, NY USA

[6] NYU Langone Hlth, Dept Mol Pharmacol & Biochem, New York, NY USA

[7] Johns Hopkins Univ, Sch Med, Dept Mol Biol & Genet, Baltimore, MD 21205 USA

[8] Johns Hopkins Univ, Sch Med, Dept Pharmacol, Baltimore, MD 21205 USA

[9] Leidos Biomed Res Inc, Canc Res Technol Program, Frederick Natl Lab Canc Res, Frederick, MD USA

[10] Johns Hopkins Univ, Sch Med, Dept Psychiat & Behav Sci, Baltimore, MD 21205 USA

[11] Johns Hopkins Univ, Sch Med, Eukaryot Tissue Culture Facil, Baltimore, MD USA

[12] NeoBiotechnologies Inc, Union City, CA USA

[13] Rutgers State Univ, Dept Mol Biol & Biochem, Piscataway, NJ USA

[14] Ctr Adv Biotechnol & Med, Piscataway, NJ USA

[15] Johns Hopkins Univ, Sch Med, Ctr Human Syst Biol, Baltimore, MD USA

[16] Johns Hopkins Univ, Sch Med, Dept Ophthalmol, Baltimore, MD 21205 USA

[17] Johns Hopkins Univ, Sch Med, Dept Neurol, Baltimore, MD 21205 USA

[18] Johns Hopkins Univ, Sch Med, Inst Cell Engn, Baltimore, MD 21205 USA

[19] Univ Texas Southwestern Med Ctr Dallas, Dept Immunol, Dallas, TX 75390 USA

[20] Sigma Aldrich Corp, BioReliance, Rockville, MD USA

来源：

NATURE METHODS | 2018年 / 15卷 / 05期

关键词：

AFFINITY; RETROTRANSPOSITION; IDENTIFICATION; SPECIFICITY; VALIDATION; EXPRESSION; PROTEINS; REAGENTS; LHX2;

D O I：

10.1038/nmeth.4632

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

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.

引用

页码：330 / +

页数：14

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