Click here to close
Hello! We notice that you are using Internet Explorer, which is not supported by Xenbase and may cause the site to display incorrectly.
We suggest using a current version of Chrome,
FireFox, or Safari.
Nat Methods
2018 May 01;155:330-338. doi: 10.1038/nmeth.4632.
Show Gene links
Show Anatomy links
A toolbox of immunoprecipitation-grade monoclonal antibodies to human transcription factors.
Venkataraman A
,
Yang K
,
Irizarry J
,
Mackiewicz M
,
Mita P
,
Kuang Z
,
Xue L
,
Ghosh D
,
Liu S
,
Ramos P
,
Hu S
,
Bayron Kain D
,
Keegan S
,
Saul R
,
Colantonio S
,
Zhang H
,
Behn FP
,
Song G
,
Albino E
,
Asencio L
,
Ramos L
,
Lugo L
,
Morell G
,
Rivera J
,
Ruiz K
,
Almodovar R
,
Nazario L
,
Murphy K
,
Vargas I
,
Rivera-Pacheco ZA
,
Rosa C
,
Vargas M
,
McDade J
,
Clark BS
,
Yoo S
,
Khambadkone SG
,
de Melo J
,
Stevanovic M
,
Jiang L
,
Li Y
,
Yap WY
,
Jones B
,
Tandon A
,
Campbell E
,
Montelione GT
,
Anderson S
,
Myers RM
,
Boeke JD
,
Fenyö D
,
Whiteley G
,
Bader JS
,
Pino I
,
Eichinger DJ
,
Zhu H
,
Blackshaw S
.
???displayArticle.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.
Abdiche,
Determining kinetics and affinities of protein interactions using a parallel real-time label-free biosensor, the Octet.
2008, Pubmed
Abdiche,
Determining kinetics and affinities of protein interactions using a parallel real-time label-free biosensor, the Octet.
2008,
Pubmed
Blackshaw,
The NIH Protein Capture Reagents Program (PCRP): a standardized protein affinity reagent toolbox.
2016,
Pubmed
Bordeaux,
Antibody validation.
2010,
Pubmed
Bradbury,
Reproducibility: Standardize antibodies used in research.
2015,
Pubmed
Chu,
Systematic discovery of Xist RNA binding proteins.
2015,
Pubmed
Cox,
Identification of SUMO E3 ligase-specific substrates using the HuProt human proteome microarray.
2015,
Pubmed
Dai,
Poly(A) binding protein C1 is essential for efficient L1 retrotransposition and affects L1 RNP formation.
2012,
Pubmed
Edgar,
Gene Expression Omnibus: NCBI gene expression and hybridization array data repository.
2002,
Pubmed
Greenspan,
Cohen's Conjecture, Howard's Hypothesis, and Ptashne's Ptruth: an exploration of the relationship between affinity and specificity.
2010,
Pubmed
Hornsby,
A High Through-put Platform for Recombinant Antibodies to Folded Proteins.
2015,
Pubmed
Hu,
Identification of new autoantigens for primary biliary cirrhosis using human proteome microarrays.
2012,
Pubmed
Hu,
DNA methylation presents distinct binding sites for human transcription factors.
2013,
Pubmed
Hu,
Profiling the human protein-DNA interactome reveals ERK2 as a transcriptional repressor of interferon signaling.
2009,
Pubmed
Jeong,
Rapid identification of monospecific monoclonal antibodies using a human proteome microarray.
2012,
Pubmed
Lee,
Tanycytes of the hypothalamic median eminence form a diet-responsive neurogenic niche.
2012,
Pubmed
Longo,
Transient mammalian cell transfection with polyethylenimine (PEI).
2013,
Pubmed
Marcon,
Assessment of a method to characterize antibody selectivity and specificity for use in immunoprecipitation.
2015,
Pubmed
McHugh,
The Xist lncRNA interacts directly with SHARP to silence transcription through HDAC3.
2015,
Pubmed
Mita,
Fluorescence ImmunoPrecipitation (FLIP): a Novel Assay for High-Throughput IP.
2016,
Pubmed
Na,
A high-throughput pipeline for the production of synthetic antibodies for analysis of ribonucleoprotein complexes.
2016,
Pubmed
Newman,
Construction of human activity-based phosphorylation networks.
2013,
Pubmed
Pauly,
How to avoid pitfalls in antibody use.
2015,
Pubmed
Rapicavoli,
The long noncoding RNA Six3OS acts in trans to regulate retinal development by modulating Six3 activity.
2011,
Pubmed
Rhodes,
Antibodies as valuable neuroscience research tools versus reagents of mass distraction.
2006,
Pubmed
Roncador,
The European antibody network's practical guide to finding and validating suitable antibodies for research.
2016,
Pubmed
Saper,
Magic peptides, magic antibodies: guidelines for appropriate controls for immunohistochemistry.
2003,
Pubmed
Schonbrunn,
Editorial: Antibody can get it right: confronting problems of antibody specificity and irreproducibility.
2014,
Pubmed
Steward,
The importance of antibody affinity in the performance of immunoassays for antibody.
1985,
Pubmed
Taylor,
Affinity proteomics reveals human host factors implicated in discrete stages of LINE-1 retrotransposition.
2013,
Pubmed
Uhlen,
A proposal for validation of antibodies.
2016,
Pubmed
Uhlén,
Proteomics. Tissue-based map of the human proteome.
2015,
Pubmed
Uzoma,
Global Identification of Small Ubiquitin-related Modifier (SUMO) Substrates Reveals Crosstalk between SUMOylation and Phosphorylation Promotes Cell Migration.
2018,
Pubmed
Vaquerizas,
A census of human transcription factors: function, expression and evolution.
2009,
Pubmed
Weller,
Quality Issues of Research Antibodies.
2016,
Pubmed
Zhu,
Global analysis of protein activities using proteome chips.
2001,
Pubmed
de Melo,
Lhx2 Is an Essential Factor for Retinal Gliogenesis and Notch Signaling.
2016,
Pubmed
de Melo,
Injury-independent induction of reactive gliosis in retina by loss of function of the LIM homeodomain transcription factor Lhx2.
2012,
Pubmed