Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S2) antibody [H5] - ChIP Grade (ab24758)

Overview

  • Product nameAnti-RNA polymerase II CTD repeat YSPTSPS (phospho S2) antibody [H5] - ChIP Grade
    See all RNA polymerase II CTD repeat YSPTSPS primary antibodies
  • Description
    Mouse monoclonal [H5] to RNA polymerase II CTD repeat YSPTSPS (phospho S2) - ChIP Grade
  • SpecificityThe antibody shows some weak reactivity towards unphosphorylated RNA polymerase II, in addition to the phosphorylated protein
  • Tested applicationsICC/IF, ChIP, WB, IPmore details
  • Species reactivity
    Reacts with: Mouse, Human, Fruit fly (Drosophila melanogaster), Zebrafish
    Predicted to work with: a wide range of other species
  • Immunogen

    Full length native protein - purified phosphorylated form of RNA polymerase II extracted from a transformed cell line.

  • General notes


    This antibody recognizes the phosphoserine 2 version of pol II.

Properties

Applications

Our Abpromise guarantee covers the use of ab24758 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
ICC/IF Use at an assay dependent concentration.
ChIP Use 2-100 µg for µg of chromatin. PubMed: 19363488
WB 1/500. Predicted molecular weight: 217 kDa.
IP 1/100.
  • Application notesIs unsuitable for IHC-FoFr.
  • 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 an 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.
    • DomainThe C-terminal domain (CTD) serves as a platform for assembly of factors that regulate transcription initiation, elongation, termination and mRNA processing.
    • Post-translational
      modifications
      The tandem heptapeptide 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 heptapeptide 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.
      Among tandem heptapeptide repeats of the C-terminal domain (CTD) some do not match the Y-S-P-T-S-P-S consensus, the seventh serine residue 'Ser-7' being replaced by a lysine. 'Lys-7' in these non-consensus heptapeptide repeats can be alternatively acetylated, methylated and dimethylated. EP300 is one of the enzyme able to acetylate 'Lys-7'. Acetylation at 'Lys-7' of non-consensus heptapeptide repeats is associated with 'Ser-2' phosphorylation and active transcription. It may regulate initiation or early elongation steps of transcription specially for inducible genes.
      Methylated at Arg-1810 prior to transcription initiation when the CTD is hypophosphorylated, phosphorylation at Ser-1805 and Ser-1808 preventing this methylation. Symmetrically or asymmetrically dimethylated at Arg-1810 by PRMT5 and CARM1 respectively. Symmetric or asymmetric dimethylation modulates interactions with CTD-binding proteins like SMN1/SMN2 and TDRD3. SMN1/SMN2 interacts preferentially with the symmetrically dimethylated form while TDRD3 interacts with the asymmetric form. Through the recruitment of SMN1/SMN2, symmetric dimethylation is required for resolving RNA-DNA hybrids created by RNA polymerase II, that form R-loop in transcription terminal regions, an important step in proper transcription termination. CTD dimethylation may also facilitate the expression of select RNAs. Among tandem heptapeptide repeats of the C-terminal domain (CTD) some do not match the Y-S-P-T-S-P-S consensus, the seventh serine residue 'Ser-7' being replaced by a lysine. 'Lys-7' in these non-consensus heptapeptide repeats can be alternatively acetylated, methylated and dimethylated. Methylation occurs in the earliest transcription stages and precedes or is concomitant to 'Ser-5' and 'Ser-7' phosphorylation.
      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.
    • Cellular localizationNucleus.
    • Information by UniProt
    • Database links
      see all
    • Alternative names
      • DNA directed RNA polymerase II A 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
      • 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
      • RPBh1 antibody
      • RpIILS antibody
      • RPO2 antibody
      • RPOL2 antibody
      see all

    Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S2) antibody [H5] - ChIP Grade images

    • ab24758 at 1/1000 dilution staining Hela cell by ICC/IF. Cells were paraformaldehyde fixed.

    • Arabidopsis thaliana root nuclei stained for RNA polymerase II with ab24758 and a polyclonal goat antimouse secondary ab conjugated to FITC (green); Nuclei are counterstained with propidium iodide (PI) - the intensively stained red regions are heterochromatic.

      See Abreview

    References for Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S2) antibody [H5] - ChIP Grade (ab24758)

    This product has been referenced in:
    • Kalendová A  et al. Nuclear actin filaments recruit cofilin and actin-related protein 3, and their formation is connected with a mitotic block. Histochem Cell Biol 142:139-52 (2014). Read more (PubMed: 25002125) »
    • Nestorov P  et al. The maternal transcriptome of the crustacean Parhyale hawaiensis is inherited asymmetrically to invariant cell lineages of the ectoderm and mesoderm. PLoS One 8:e56049 (2013). ICC/IF ; Parhyale hawaiensis . Read more (PubMed: 23418507) »

    See all 26 Publications for this product

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    ab5408 is part of the RNA polymerase II CTD Panel reference ab103968 which is composed of:

    25µg of anti-RNA polymerase II CTD repeat YSPTSPS (phospho S2) antibody - ChIP Grade (http://www.abcam.com/a...

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    I can confirm that the isotype for the H5 RNA polymerase antibody is IgM as indicated on the datasheet.

    Abcam has not validated the combination of species/application used in this Abreview.
    Application ELISA
    Sample Human Recombinant protein (RNA polymerase II CTD repead YSPTSPS)
    Specification RNA polymerase II CTD repead YSPTSPS
    Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 0.1% · Temperature: 22°C
    Type Indirect
    Username

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    Verified customer

    Submitted Feb 21 2013

    Many thanks for contacting us and your interest in out products.

    The RNA polymerase II protein has a protein sequence which predicts its size to be 217 kDa (SwissProt reference P24928). This is where the figure on the datasheet of the Anti-RNA...

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    Le numéro de commande pour ab26721 et ab1791 est xxxxxxxx. Vous recevrez prochainement un mail de confirmation comprenant les détails d'expédition.

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    J’ai eu ma réponse de nos laboratoires et ce cas et assez compliqué. Premièrement je tiens à vous remercier d’avoir patienté et j’apprécie &eacu...

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    I am happy to see that the image shows a clear band in the first couple of samples at approximately the right size.

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