Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S5) antibody [8A7] (ab5401)

Overview

  • Product nameAnti-RNA polymerase II CTD repeat YSPTSPS (phospho S5) antibody [8A7]See all RNA polymerase II CTD repeat YSPTSPS primary antibodies ...
  • Description
    Mouse monoclonal [8A7] to RNA polymerase II CTD repeat YSPTSPS (phospho S5)
  • Tested applicationsFlow Cyt, ICC/IF, ELISA, WBmore details
  • Species reactivity
    Reacts with: Human, Saccharomyces cerevisiae
    Predicted to work with: Mouse, Hamster, Arabidopsis thaliana, Fruit fly (Drosophila melanogaster), Schizosaccharomyces pombe
  • Immunogen

    Synthetic peptide: 10 repeats of YSPTSpPS (Human).

Properties

Applications

Our Abpromise guarantee covers the use of ab5401 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
Flow Cyt Use 1µg for 106 cells.
ICC/IF Use at an assay dependent concentration.
ELISA Use at an assay dependent dilution.
WB Use at an assay dependent dilution. Predicted molecular weight: 217 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
    • 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 pol II CTD 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 S5) antibody [8A7] images

  • ab5401 staining RNA polymerase II CTD repeat YSPTSPS (phospho S5) in Human HeLa cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with methanol and blocked with 0.2% Fish scale gelatin for 20 minutes at 25°C. Samples were incubated with primary antibody (1/500 PBS + 0.2% fish scale gelatin) for 20 minutes at 25°C. An Alexa Fluor®594-conjugated Donkey anti-mouse IgG polyclonal (1/500) was used as the secondary antibody.
    Left-hand panel: DAPI nuclear staining.
    Centre-hand panel: ab5401 staining (red).
    Right-hand panel: Merged

    See Abreview

  • Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S5) antibody [8A7] (ab5401) at 1/3000 dilution + whole cell lysate prepared from murine stem cells at 100000 cells

    Secondary
    HRP conjugated goat anti-mouse IgG (H+L) at 1/4000 dilution
    developed using the ECL technique

    Predicted band size : 217 kDa


    Exposure time : 1 minute

    Image courtesy of an anonymous Abreview.

    See Abreview

  • Overlay histogram showing HeLa cells stained with ab5401 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab5401, 1µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.

References for Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S5) antibody [8A7] (ab5401)

This product has been referenced in:
  • Bolt MJ  et al. Systems level-based RNAi screening by high content analysis identifies UBR5 as a regulator of estrogen receptor-a protein levels and activity. Oncogene N/A:N/A (2014). Human . Read more (PubMed: 24441042) »
  • Chang PV  et al. The microbial metabolite butyrate regulates intestinal macrophage function via histone deacetylase inhibition. Proc Natl Acad Sci U S A 111:2247-52 (2014). Read more (PubMed: 24390544) »

See all 3 Publications for this product

Product Wall

Abcam guarantees this product to work in the 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
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Verified customer

Submitted Feb 21 2013

Application Western blot
Sample Mouse Cell lysate - whole cell (Epiblast stem cells)
Loading amount 100000 cells
Specification Epiblast stem cells
Gel Running Conditions Non-reduced Denaturing
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
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Submitted Jan 19 2012

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunocytochemistry/ Immunofluorescence
Sample Human Cell (Hela cell)
Specification Hela cell
Fixative Methanol
Permeabilization No
Blocking step Fish scale gelatin as blocking agent for 20 minute(s) · Concentration: 0.2% · Temperature: 25°C
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Verified customer

Submitted Jun 07 2010

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Human Cell lysate - whole cell (MCF7 breast carcinoma cell line)
Loading amount 30 µg
Specification MCF7 breast carcinoma cell line
Treatment DMSO or CAN508 for 24hr
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5%
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Verified customer

Submitted Jan 02 2007

Thank you for your enquiry. Unfortunately, we regret to inform you that there is no Western blot image available. May I ask you if you have already purchased this antibody or just planning to buy it? This item is currently out of stock. The ...

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