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    rna-polymerase-ii-ctd-repeat-ysptsps-phospho-t4-antibody-6d7-ab252851.pdf

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Epigenetics and Nuclear Signaling Transcription Polymerase associated factors Pol II Transcription
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Recombinant

Recombinant Anti-RNA polymerase II CTD repeat YSPTSPS (phospho T4) antibody [6D7] (ab252851)

  • Datasheet
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Western blot - Anti-RNA polymerase II CTD repeat YSPTSPS (phospho T4) antibody [6D7] (ab252851)
  • Indirect ELISA - Anti-RNA polymerase II CTD repeat YSPTSPS (phospho T4) antibody [6D7] (ab252851)
  • Dot Blot - Anti-RNA polymerase II CTD repeat YSPTSPS (phospho T4) antibody [6D7] (ab252851)
  • Anti-RNA polymerase II CTD repeat YSPTSPS (phospho T4) antibody [6D7] (ab252851)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rat monoclonal [6D7] to RNA polymerase II CTD repeat YSPTSPS (phospho T4)
  • Suitable for: Indirect ELISA, WB, Dot blot
  • Reacts with: Rat, Human

Conjugates logo Related conjugates and formulations

Carrier Free

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Overview

  • Product name

    Anti-RNA polymerase II CTD repeat YSPTSPS (phospho T4) antibody [6D7]
    See all RNA polymerase II CTD repeat YSPTSPS primary antibodies
  • Description

    Rat monoclonal [6D7] to RNA polymerase II CTD repeat YSPTSPS (phospho T4)
  • Host species

    Rat
  • Tested applications

    Suitable for: Indirect ELISA, WB, Dot blotmore details
    Unsuitable for: ChIP or ICC
  • Species reactivity

    Reacts with: Rat, Human
  • Immunogen

    The details of the immunogen for this antibody are not available.

  • Positive control

    • WB: PC-12 whole cell lysate.
  • General notes

    This antibody clone is manufactured by Abcam. If you require a custom buffer formulation or conjugation for your experiments, please contact orders@abcam.com.

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
  • Storage buffer

    Preservative: 0.01% Sodium azide
    Constituents: 0.05% BSA, 40% Glycerol (glycerin, glycerine), 59.94% PBS
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    6D7
  • Isotype

    IgG2b
  • Research areas

    • Epigenetics and Nuclear Signaling
    • Transcription
    • Polymerase associated factors
    • Pol II Transcription
    • Epigenetics and Nuclear Signaling
    • Transcription
    • Polymerase associated factors
    • Pol II Transcription
    • Polymerase
    • Epigenetics and Nuclear Signaling
    • Transcription
    • RNA polymerase

Associated products

  • Alternative Versions

    • Anti-RNA polymerase II CTD repeat YSPTSPS (phospho T4) antibody [6D7] - BSA and Azide free (ab255845)
  • Compatible Secondaries

    • Goat Anti-Rat IgG H&L (HRP) (ab205720)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab252851 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
Indirect ELISA
Use at an assay dependent concentration.

Use at 125 ng/ml.

WB
1/1000. Detects a band of approximately 260 kDa (predicted molecular weight: 217 kDa).
Dot blot
1/1000.
Notes
Indirect ELISA
Use at an assay dependent concentration.

Use at 125 ng/ml.

WB
1/1000. Detects a band of approximately 260 kDa (predicted molecular weight: 217 kDa).
Dot blot
1/1000.
Application notes
Is unsuitable for ChIP or ICC.

Target

  • Function

    DNA-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 similarities

    Belongs to the RNA polymerase beta' chain family.
  • Domain

    The 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 localization

    Nucleus.
  • 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: 5430 Human
    • Entrez Gene: 363633 Rat
    • Omim: 180660 Human
    • SwissProt: P24928 Human
    • Unigene: 270017 Human
    • 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

    Images

    • Western blot - Anti-RNA polymerase II CTD repeat YSPTSPS (phospho T4) antibody [6D7] (ab252851)
      Western blot - Anti-RNA polymerase II CTD repeat YSPTSPS (phospho T4) antibody [6D7] (ab252851)
      All lanes : Anti-RNA polymerase II CTD repeat YSPTSPS (phospho T4) antibody [6D7] (ab252851) at 1/1000 dilution

      Lane 1 : PC-12 (rat adrenal gland pheochromocytoma), whole cell lysate (Untreated membrane)
      Lane 2 : PC-12 whole cell lysate (Phosphatase treated membrane)

      Lysates/proteins at 10 µg per lane.

      Secondary
      All lanes : Goat Anti-Rat IgG H&L (HRP) (ab205720) at 1/5000 dilution

      Predicted band size: 217 kDa
      Observed band size: 260 kDa why is the actual band size different from the predicted?


      Exposure time: 3 minutes


      The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID: 22745433 and 23071310).

      This blot was developed using a higher sensitivity ECL substrate.
      This antibody reacts with rat on WB.

      Blocking/Dilution buffer: 5% NFDM/TBST.

    • Indirect ELISA - Anti-RNA polymerase II CTD repeat YSPTSPS (phospho T4) antibody [6D7] (ab252851)
      Indirect ELISA - Anti-RNA polymerase II CTD repeat YSPTSPS (phospho T4) antibody [6D7] (ab252851)

      ab252851 used at 0-1000 ng/ml.

      Antigen (phospho S2, phospho S5, phospho T4, phospho Y1, phospho S7, non-phospho) used at 500 ng/ml.

      Secondary antibody: Alkaline Phosphatase-conjugated AffiniPure Goat Anti-Rat IgG (H+L) used at a 1/1000 dilution.

    • Dot Blot - Anti-RNA polymerase II CTD repeat YSPTSPS (phospho T4) antibody [6D7] (ab252851)
      Dot Blot - Anti-RNA polymerase II CTD repeat YSPTSPS (phospho T4) antibody [6D7] (ab252851)

      ab252851 used at 1/1000 dilution, followed by Goat Anti-Rat IgG (H+L), HRP) (ab205720) at 1/5000 dilution.

      Lane 1: RNA polymerase II CTD repeat YSPTSPS (phospho S2) peptide

      Lane 2: RNA polymerase II CTD repeat YSPTSPS (phospho S5) peptide

      Lane 3: RNA polymerase II CTD repeat YSPTSPS (phospho T4) peptide

      Lane 4: RNA polymerase II CTD repeat YSPTSPS (phospho Y1) peptide

      Lane 5: RNA polymerase II CTD repeat YSPTSPS (phospho S7) peptide

      Lane 6: RNA polymerase II CTD repeat YSPTSPS (non-phospho) peptide

      Blcoking/Dilution buffer: 5% NFDM/TBST.

      Exposure time 3 mins.

    • Anti-RNA polymerase II CTD repeat YSPTSPS (phospho T4) antibody [6D7] (ab252851)
      Anti-RNA polymerase II CTD repeat YSPTSPS (phospho T4) antibody [6D7] (ab252851)

    Protocols

    To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

    Click here to view the general protocols

    Datasheets and documents

    • Datasheet download

      Download

    References (1)

    Publishing research using ab252851? Please let us know so that we can cite the reference in this datasheet.

    ab252851 has been referenced in 1 publication.

    • Hintermair C  et al. Threonine-4 of mammalian RNA polymerase II CTD is targeted by Polo-like kinase 3 and required for transcriptional elongation. EMBO J 31:2784-97 (2012). PubMed: 22549466

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