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, WB more 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 YSPTS_pPS (Human).

  Run BLAST with Run BLAST with

Properties

• FormLiquid
• Storage instructionsShipped at 4°C. Upon delivery aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
• Storage bufferPreservative: None
  Constituents: PBS
• Concentration information loading...
• PurityProtein A purified
• Clonality Monoclonal
• Clone number8A7
• MyelomaSp2/0-Ag14
• IsotypeIgG1
• Research Areas
  • Epigenetics and Nuclear Signaling
  • Transcription
  • Polymerase associated factors
  • Pol II Transcription
  • Polymerase

  • Epigenetics and Nuclear Signaling
  • Transcription
  • RNA polymerase

Applications

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
  modificationsThe 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.

Target information above from: UniProt accession P24928 The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010) .

Information by UniProt
• Database links
  • Entrez Gene: 32100 Fruit fly (Drosophila melanogaster)
  • Entrez Gene: 5430 Human
  • Entrez Gene: 20020 Mouse
  • Entrez Gene: 851415 Saccharomyces cerevisiae
  • Omim: 180660 Human
  • SwissProt: P18616 Arabidopsis thaliana
  • SwissProt: P04052 Fruit fly (Drosophila melanogaster)
  • SwissProt: P24928 Human

  • SwissProt: P08775 Mouse
  • Unigene: 2925 Fruit fly (Drosophila melanogaster)
  • Unigene: 270017 Human
  • Unigene: 16533 Mouse

  see all

• Alternative names
  DNA directed RNA polymerase II A antibodyDNA-directed RNA polymerase II largest subunit antibodyDNA-directed RNA polymerase II largest subunit RNA polymerase II 220 kd subunit antibody

  DNA-directed RNA polymerase II subunit A antibodyDNA-directed RNA polymerase II subunit RPB1 antibodyDNA-directed RNA polymerase III largest subunit antibodyhRPB220 antibodyhsRPB1 antibodyPOLR2 antibodyPolr2a antibodyPOLRA antibodyPolymerase (RNA) II (DNA directed) polypeptide A 220kDa antibodyPolymerase (RNA) II (DNA directed) polypeptide A antibodyRNA pol II CTD antibodyRNA polymerase II subunit B1 antibodyRNA-directed RNA polymerase II subunit RPB1 antibodyRPB1 antibodyRPB1_HUMAN antibodyRPBh1 antibodyRpIILS antibodyRPO2 antibodyRPOL2 antibody

  see all

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

• 

  Immunocytochemistry/ Immunofluorescence - RNA polymerase II CTD repeat YSPTSPS (phospho S5) antibody [8A7] (ab5401)This image is courtesy of an anonymous Abreview
  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

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• 

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

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• 

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