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Monoclonal Antibodies

Anti-Phospho RNA polII (Ser5) [CTD 8A7]

Invented by Dr Jesper Svejstrup from Francis Crick Institute
Invented at Cancer Research UK London Research Institute: Clare Hall Laboratories
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Monoclonal antibody 8A7. RNA polymerase II (RNAP II) contains 12 subunits. The C-terminal domain of the largest subunit of RNAP II is reversibly phosphorylated on the highly conserved heptapeptide repeat, YSPTSPS. The major physiological phosphorylation sites are Ser-2 and Ser-5. Monoclonal antibody 8A7 was obtained using synthetic phosphopeptides and specifically recognises the Ser-5 phosphorylated form of RNAP II.
Adapted from : Szafran AT, Stossi F, Mancini MG, Walker CL, Mancini MA (2017) Characterizing properties of non-estrogenic substituted bisphenol analogs using high throughput microscopy and image analysis. PLoS ONE 12(7): e0180141 PRL array targeting of phospho-serine 5 RNA polymeraseII (pS5-RNAPII) following 1 hr of compound treatment in the presence of 10 nM E2, assayed by immunofluorescence.
Adapted from Treviño et al. Mol Endocrinol, February 2016, 30(2):158 –172 High-throughput microscopy analysis of the effect of the agonist R5020 and antagonists, RU486 and ZK98299, on PR transcriptional activity. Quantification of the effect of RU486 on hormone-mediated SRC-1, SRC-3, and MED14 coregulator protein recruitment to the array, as well as phosphorylation of RNA Pol II on serine 5 (an indicator of activated RNA Pol II) at the array. Cells were pretreated with RU486, treated with R5020 as indicated in C, and then processed for immunofluorescence with antibodies against the proteins of interest. Graph represents the combined data from 3 independent experiments. Relative protein recruitment (yaxis) is the indirect immunofluorescence intensity as a result of coactivator-specific antibody binding to the coactivator bound at the array. *, P <.05; **, P < .01; ***, P < .001.
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Catalogue Number 151243
Applications ELISA FACS IF WB
Antigen/Gene or Protein Targets RNA Polymerase II (Phosphorylated)
Reactivity Human and Saccharomyces cerevisiae
Relevance RNAPII is an essential component of the RNAP II transcription elongation complex. Recruitment of RNA processing factors to the elongation complex is coordinated by the phosphorylation of Serine-2 and Serine-5 residues within the C-terminal repeat (YSPTSPS) domain of the largest subunit of RNAPII.
Host Mouse
Immunogen Peptide: 10 repeats of synthetic peptide YSPTSPS using chemically sythesized phospho-Ser5.
Positive Control S. cerevisiae and HeLa cells
Subclass IgM
Myeloma Used Sp2/0-Ag14
Recommended Growing Conditions DMEM + 5% FCS
Strain Balb/c
Research Area Epigenetics & Nuclear Signalling

References

There are 10 reference entries for this reagent.

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References: 10 entries

Szafran et al. 2017. PLoS One. 12(7):e0180141. PMID: 28704378.

IF

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Treviño et al. 2016. Mol Endocrinol. 30(2):158-72. PMID: 26652902.

IF

View  

Benferhat et al. 2012. J Virol. 86(20):11333-44. PMID: 22896612.

IF

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Large-scale chromatin immunoprecipitation with promoter sequence microarray analysis of the interaction of the NSs protein of Rift Valley fever virus with regulatory DNA regions of the host genome.

Europe PMC ID: 22896612

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Alm-Kristiansen et al. 2011. Mol Cancer. 10:21. PMID: 21338522.

IF

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PIAS1 interacts with FLASH and enhances its co-activation of c-Myb.

Europe PMC ID: 21338522

View  

Aune et al. 2008. Clin Cancer Res. 14(20):6449-55. PMID: 18927284.

WB

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Von Hippel-Lindau-coupled and transcription-coupled nucleotide excision repair-dependent degradation of RNA polymerase II in response to trabectedin.

Europe PMC ID: 18927284

View  

Otero et al. 1999. Mol Cell. 3(1):109-18. PMID: 10024884.

View  

Elongator, a multisubunit component of a novel RNA polymerase II holoenzyme for transcriptional elongation.

Europe PMC ID: 10024884

View  

Add a reference

References: 10 entries

Szafran et al. 2017. PLoS One. 12(7):e0180141. PMID: 28704378.

IF

View  

Treviño et al. 2016. Mol Endocrinol. 30(2):158-72. PMID: 26652902.

IF

View  

Benferhat et al. 2012. J Virol. 86(20):11333-44. PMID: 22896612.

IF

View  

Large-scale chromatin immunoprecipitation with promoter sequence microarray analysis of the interaction of the NSs protein of Rift Valley fever virus with regulatory DNA regions of the host genome.

View  

Alm-Kristiansen et al. 2011. Mol Cancer. 10:21. PMID: 21338522.

IF

View  

PIAS1 interacts with FLASH and enhances its co-activation of c-Myb.

View  

Aune et al. 2008. Clin Cancer Res. 14(20):6449-55. PMID: 18927284.

WB

View  

Von Hippel-Lindau-coupled and transcription-coupled nucleotide excision repair-dependent degradation of RNA polymerase II in response to trabectedin.

View  

Otero et al. 1999. Mol Cell. 3(1):109-18. PMID: 10024884.

View  

Elongator, a multisubunit component of a novel RNA polymerase II holoenzyme for transcriptional elongation.

View  

Add a reference

Inventor Information