Anti-RNA polymerase II CTD repeat YSPTSPS antibody [H14] - ChIP Grade (ab24759)

Overview

  • Product nameAnti-RNA polymerase II CTD repeat YSPTSPS antibody [H14] - ChIP GradeSee all RNA polymerase II CTD repeat YSPTSPS primary antibodies ...
  • Description
    Mouse monoclonal [H14] to RNA polymerase II CTD repeat YSPTSPS - ChIP Grade
  • SpecificityThis antibody recognizes the phosphoserine 5 version of pol II. The antibody also shows some weak reactivity towards unphosphorylated RNA polymerase II, in addition to the phosphorylated protein
  • Tested applicationsWB, ChIP, ICC, ICC/IFmore details
  • Species reactivity
    Reacts with: Mouse, Human, Saccharomyces cerevisiae, Schizosaccharomyces pombe
    Predicted to work with: a wide range of other species
  • Immunogen

    Phosphorylated form of RNA polymerase II purified from a transformed cell line.

  • General notesFor western blots use casein or BSA in the blocking solution rather than non-fat-dry milk.

Properties

Applications

Our Abpromise guarantee covers the use of ab24759 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
WB 1/500. Predicted molecular weight: 220-240 kDa. This antibody recognizes the phosphoserine 5 version of pol II. The antibody also shows some weak reactivity towards unphosphorylated RNA polymerase II, in addition to the phosphorylated protein.
ChIP Use at an assay dependent concentration. PubMed: 21880597
ICC Use at an assay dependent concentration.
ICC/IF Use at an assay dependent concentration.

Target

  • FunctionDNA-dependent RNA polymerase catalyzes the transcription of DNA into RNA using the four ribonucleoside triphosphates as substrates. Largest and catalytic component of RNA polymerase II which synthesizes mRNA precursors and many functional non-coding RNAs. Forms the polymerase active center together with the second largest subunit. Pol II is the central component of the basal RNA polymerase II transcription machinery. It is composed of mobile elements that move relative to each other. RPB1 is part of the core element with the central large cleft, the clamp element that moves to open and close the cleft and the jaws that are thought to grab the incoming DNA template. At the start of transcription, a single-stranded DNA template strand of the promoter is positioned within the central active site cleft of Pol II. A bridging helix emanates from RPB1 and crosses the cleft near the catalytic site and is thought to promote translocation of Pol II by acting as a ratchet that moves the RNA-DNA hybrid through the active site by switching from straight to bent conformations at each step of nucleotide addition. During transcription elongation, Pol II moves on the template as the transcript elongates. Elongation is influenced by the phosphorylation status of the C-terminal domain (CTD) of Pol II largest subunit (RPB1), which serves as a platform for assembly of factors that regulate transcription initiation, elongation, termination and mRNA processing. Acts as a RNA-dependent RNA polymerase when associated with small delta antigen of Hepatitis delta virus, acting both as a replicate and transcriptase for the viral RNA circular genome.
  • Sequence similaritiesBelongs to the RNA polymerase beta' chain family.
  • Post-translational
    modifications
    The tandem 7 residues repeats in the C-terminal domain (CTD) can be highly phosphorylated. The phosphorylation activates Pol II. Phosphorylation occurs mainly at residues 'Ser-2' and 'Ser-5' of the heptapeptide repeat and is mediated, at least, by CDK7 and CDK9. CDK7 phosphorylation of POLR2A associated with DNA promotes transcription initiation by triggering dissociation from DNA. Phosphorylation also takes place at 'Ser-7' of the heptapepdtide repeat, which is required for efficient transcription of snRNA genes and processing of the transcripts. The phosphorylation state is believed to result from the balanced action of site-specific CTD kinases and phosphatases, and a 'CTD code' that specifies the position of Pol II within the transcription cycle has been proposed.
    Dephosphorylated by the protein phosphatase CTDSP1.
    Ubiquitinated by WWP2 leading to proteasomal degradation (By similarity). Following UV treatment, the elongating form of RNA polymerase II (RNA pol IIo) is ubiquitinated UV damage sites without leading to degradation: ubiquitination is facilitated by KIAA1530/UVSSA and promotes RNA pol IIo backtracking to allow access to the nucleotide excision repair machinery.
    Methylated at Arg-1810 by CARM1. Methylation occurs only when the CTD is hypophosphorylated, and phosphorylation at Ser-1805 and Ser-1808 prevent methylation (in vitro). It is assumed that methylation occurs prior to phosphorylation and transcription initiation. CTD methylation may facilitate the expression of select RNAs.
  • Cellular localizationNucleus.
  • Information by UniProt
  • Database links
  • Alternative names
    • B220 antibody
    • DNA directed RNA polymerase II A antibody
    • DNA-directed RNA polymerase II largest subunit antibody
    • DNA-directed RNA polymerase II largest subunit RNA polymerase II 220 kd subunit antibody
    • DNA-directed RNA polymerase II subunit A antibody
    • DNA-directed RNA polymerase II subunit RPB1 antibody
    • DNA-directed RNA polymerase III largest subunit antibody
    • hRPB220 antibody
    • hsRPB1 antibody
    • MGC75453 antibody
    • POLR2 antibody
    • POLR2A antibody
    • POLRA antibody
    • Polymerase (RNA) II (DNA directed) polypeptide A 220kDa antibody
    • Polymerase (RNA) II (DNA directed) polypeptide A antibody
    • RNA polymerase II subunit B1 antibody
    • RNA-directed RNA polymerase II subunit RPB1 antibody
    • RPB1 antibody
    • RPB1_HUMAN antibody
    • RPB220 antibody
    • RPBh1 antibody
    • RpIILS antibody
    • RPO2 antibody
    • RPOL2 antibody
    see all

References for Anti-RNA polymerase II CTD repeat YSPTSPS antibody [H14] - ChIP Grade (ab24759)

This product has been referenced in:
  • Tao Y  et al. Treatment of breast cancer cells with DNA demethylating agents leads to a release of Pol II stalling at genes with DNA-hypermethylated regions upstream of TSS. Nucleic Acids Res : (2011). ChIP ; Human . Read more (PubMed: 21880597) »
  • Tao Y  et al. Lsh Mediated RNA Polymerase II Stalling at HoxC6 and HoxC8 Involves DNA Methylation. PLoS One 5:e9163 (2010). WB ; Mouse . Read more (PubMed: 20161795) »

See all 7 Publications for this product

Product Wall

Thank you for contacting us. Because we carry over 90,000 products, it isn't feasible for us to keep small sample sizes of our products.

We are happy to reassure our customers that all of our products are covered by our Abpromise, which guaran...

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Thank you for contacting us. Although as you say the IgM is not bound well by protein A or G this antibody has been used previously with Protein G and Protein A/G beads successfully for immunoprecipitation for performing ChIP: Protein G beads: ...

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Abcam has not validated the combination of species/application used in this Abreview.
Application Immunohistochemistry (Frozen sections)
Sample Rat Tissue sections (Brain)
Specification Brain
Fixative Paraformaldehyde
Permeabilization No
Username

Dr. Karine Thibault

Verified customer

Submitted May 14 2009

Application Immunocytochemistry
Sample Human Cell (human adenocarcinoma HT29 cells)
Specification human adenocarcinoma HT29 cells
Fixative Paraformaldehyde
Permeabilization Yes - Triton X100 and Saponin
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 37°C
Username

Dr. Eva Bartova

Verified customer

Submitted Apr 01 2008

Thank you for your telephone enquiry yesterday. I have investigated this further and can confirm that both these RNA POL II antibodies (ab24758 and ab24759) have been tested to detect mouse RNA POL II I have included a reference below that uses s...

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Application Immunocytochemistry/ Immunofluorescence
Sample Human Cultured Cells (colon adenocarcinoma HT29 cells)
Specification colon adenocarcinoma HT29 cells
Fixative Paraformaldehyde
Permeabilization Yes - Triton X100 and Saponin
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: RT°C
Username

Dr. Eva Bartova

Verified customer

Submitted Nov 21 2007

Application ChIP
Sample Saccharomyces cerevisiae Cell lysate - whole cell (yeast)
Specification yeast
Type Cross-linking (X-ChIP)
Duration of cross-linking step: 2 minute(s) and 30 second(s)
Specification of the cross-linking agent: Formaldehyde
Detection step Real-time PCR
Positive control Pol II
Histone H3
Negative control Mouse IgG
Beads only
Username

Abcam user community

Verified customer

Submitted Jul 18 2007

Application Immunocytochemistry/ Immunofluorescence
Sample Human Cell (Hela)
Specification Hela
Fixative Paraformaldehyde
Blocking step BSA as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 2
Username

Abcam user community

Verified customer

Submitted Jul 11 2007

Thank you for your enquiry. I apologise for the delay in responding to your message. It has taken some time to obtain some information. I can confirm that when these H5 and H14 antibodies were first used, they detected a protein initially named...

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Application Immunocytochemistry/ Immunofluorescence
Sample Mouse Cell (testicular germ cells)
Specification testicular germ cells
Fixative Paraformaldehyde
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 0.5
Username

Dr. Claudio Sette

Verified customer

Submitted Jan 23 2007

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"