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 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
      see all
    • 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
      • 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

    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:
    • 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) »
    • Kosters A  et al. Sexually dimorphic genome-wide binding of retinoid X receptor alpha (RXRa) determines male-female differences in the expression of hepatic lipid processing genes in mice. PLoS One 8:e71538 (2013). CHIPseq ; Mouse . Read more (PubMed: 23977068) »

    See all 23 Publications for this product

    Product Wall


    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

    Abcam user community

    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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    Merci de nous avoir contactés.

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

    Le numéro de com...

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    Nous sommes désolés d'apprendre que les produits que vous avez reçus ne fonctionnent pas comme attendu. Je me permets de vous transférer le numéro de commande des rempl...

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    Nous sommes désolés d'apprendre que les produits que vous avez reçus ne fonctionnent pas comme attendu. Je suis d’accord pour vous envoyer ab26721, ab106293 et ab1791 en rempla...

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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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    Thank you for contacting us.

    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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    Thank you for your reply and for sending an image.

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

    I can suggest to use the following protocol for transfering big p...

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    Thank you for taking the time to complete our questionnaire and contact us. I am sorry to hear you have had difficulty obtaining satisfactory results from this antibody.

    The details you have kindly provided will enable us to investigate this...

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