Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S2) antibody (ab70324)

Overview

  • Product nameAnti-RNA polymerase II CTD repeat YSPTSPS (phospho S2) antibody
    See all RNA polymerase II CTD repeat YSPTSPS primary antibodies
  • Description
    Rabbit polyclonal to RNA polymerase II CTD repeat YSPTSPS (phospho S2)
  • Tested applicationsIHC-P, WBmore details
  • Species reactivity
    Reacts with: Mouse, Human
    Predicted to work with: Rat, Rabbit, Cow, Dog, Pig, Chimpanzee, Fruit fly (Drosophila melanogaster), Plants, Zebrafish, Rhesus monkey, Gorilla, Orangutan, Xenopus tropicalis
  • Immunogen

    A phosphorylated synthetic peptide of human RNA Polymerase II, from around serine 2 of the C-terminal repeated sequence YSPTSPS.

  • Positive control
    • Whole cell lysate from HeLa cells

Properties

Applications

Our Abpromise guarantee covers the use of ab70324 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
IHC-P 1/200 - 1/1000. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
WB 1/2500 - 1/20000. Detects a band of approximately 270 kDa.

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
    • 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

Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S2) antibody images

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human ovarian carcinoma mock phosphatase treated section (left) and calf intestinal phosphatase-treated section (right) tissues labelling RNA polymerase II CTD repeat YSPTSPS (phospho S2) with ab70324 at 1/500 (0.4µg/ml). Detection: DAB.
  • All lanes : Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S2) antibody (ab70324) at 0.02 µg/ml

    Lane 1 : Whole cell lysate from HeLa cells with no peptide
    Lane 2 : Whole cell lysate from HeLa cells with non-phosphorylated peptide
    Lane 3 : Whole cell lysate from HeLa cells with phosphorylated S2 peptide
    Lane 4 : Whole cell lysate from HeLa cells with phosphorylated S5 peptide

    Lysates/proteins at 50 µg per lane.


    Observed band size : 270 kDa (why is the actual band size different from the predicted?)

References for Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S2) antibody (ab70324)

ab70324 has not yet been referenced specifically in any publications.

Product Wall

Thank you for contacting Abcam and thank you for your patience while I consulted my coleagues about your enquiry. Below is the response that I received from a colleague: "I would suggest using the non phosphorylated form of Pol2 as a positive control f...

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Please accept my apologies for misunderstanding your original email. I have to admit that I am no expert in ChIP assays but I have sent an email to ask my colleagues and I will let you know what they say. I see no reason why you could not use S...

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Thank you for contacting Abcam about ab70324. As long as there is a sufficient concentration of your sample potein present, the anti-RNA polymerase II CTD repeat YSPTSPS (phospho S2) antibody should work. As it has not been tested in ChIP, we...

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