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    rna-polymerase-ii-ctd-repeat-ysptsps-phospho-s5-antibody-8a7-ab5401.pdf

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Epigenetics and Nuclear Signaling Transcription Polymerase associated factors Pol II Transcription
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Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S5) antibody [8A7] (ab5401)

  • Datasheet
Reviews (5)Q&A (1)References (9)

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

    Key features and details

    • Mouse monoclonal [8A7] to RNA polymerase II CTD repeat YSPTSPS (phospho S5)
    • Suitable for: Flow Cyt
    • Reacts with: Human
    • Isotype: IgG1

    You may also be interested in

    Primary
    Product image
    Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S2) antibody [EPR18855] (ab193468)
    Secondary
    Product image
    Goat Anti-Mouse IgG H&L (HRP) (ab205719)
    Protein
    Recombinant human RNA polymerase II CTD repeat YSPTSPS protein (ab81888)

    View more associated products

    Overview

    • Product name

      Anti-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)
    • Host species

      Mouse
    • Tested applications

      Suitable for: Flow Cytmore details
    • Species reactivity

      Reacts with: Human
      Predicted to work with: Mouse
    • Immunogen

      Synthetic peptide corresponding to Human RNA polymerase II CTD repeat YSPTSPS (phospho S5). Synthetic peptide: 10 repeats of YSPTSpPS (Human).
      Sequence: YSPTSpPS
      Database link: P24928

      Run BLAST with BLAST the sequence with ExPASy Run BLAST with BLAST the sequence with NCBI
    • General notes

      The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.

      If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As

    Properties

    • Form

      Liquid
    • Storage instructions

      Shipped at 4°C. Upon delivery aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
    • Storage buffer

      Constituent: PBS
    • Concentration information loading...
    • Purity

      Protein A purified
    • Clonality

      Monoclonal
    • Clone number

      8A7
    • Myeloma

      Sp2/0-Ag14
    • Isotype

      IgG1
    • 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

    • Compatible Secondaries

      • Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) (ab150113)
      • Goat Anti-Mouse IgG H&L (HRP) (ab205719)
      • Goat Anti-Mouse IgG H&L (DyLight® 488) preadsorbed (ab96879)
    • Isotype control

      • Mouse IgG1, kappa monoclonal [15-6E10A7] - Isotype Control (ab170190)
      • Mouse IgG1, Kappa Monoclonal [B11/6] - Isotype Control (ab91353)
    • Recombinant Protein

      • Recombinant human RNA polymerase II CTD repeat YSPTSPS protein (ab81888)

    Applications

    The Abpromise guarantee

    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.

    ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.

     

    Notes
    Flow Cyt
    Use 1µg for 106 cells.

    ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.

     

    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: 20020 Mouse
      • Omim: 180660 Human
      • SwissProt: P24928 Human
      • SwissProt: P08775 Mouse
      • Unigene: 270017 Human
      • Unigene: 16533 Mouse
      • 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

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

        Overlay histogram showing HeLa (Human epithelial cell line from cervix adenocarcinoma) 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.

      Protocols

      • Flow cytometry protocols

      Click here to view the general protocols

      Datasheets and documents

      • Datasheet download

        Download

      References (9)

      Publishing research using ab5401? Please let us know so that we can cite the reference in this datasheet.

      ab5401 has been referenced in 9 publications.

      • Szafran AT  et al. Characterizing properties of non-estrogenic substituted bisphenol analogs using high throughput microscopy and image analysis. PLoS One 12:e0180141 (2017). PubMed: 28704378
      • Meuris F  et al. Symptomatic Improvement in Human Papillomavirus-Induced Epithelial Neoplasia by Specific Targeting of the CXCR4 Chemokine Receptor. J Invest Dermatol 136:473-80 (2016). PubMed: 26967480
      • Treviño LS  et al. Differential Regulation of Progesterone Receptor-Mediated Transcription by CDK2 and DNA-PK. Mol Endocrinol 30:158-72 (2016). PubMed: 26652902
      • 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 . 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). PubMed: 24390544
      • Stossi F  et al. Defining estrogenic mechanisms of bisphenol A analogs through high throughput microscopy-based contextual assays. Chem Biol 21:743-53 (2014). PubMed: 24856822
      • Bolt MJ  et al. Coactivators enable glucocorticoid receptor recruitment to fine-tune estrogen receptor transcriptional responses. Nucleic Acids Res 41:4036-48 (2013). ICC . PubMed: 23444138
      • Krystof V  et al. 4-arylazo-3,5-diamino-1H-pyrazole CDK inhibitors: SAR study, crystal structure in complex with CDK2, selectivity, and cellular effects. J Med Chem 49:6500-9 (2006). WB ; Human . PubMed: 17064068
      • Otero G  et al. Elongator, a multisubunit component of a novel RNA polymerase II holoenzyme for transcriptional elongation. Mol Cell 3:109-18 (1999). PubMed: 10024884

      Customer reviews and Q&As

      Show All Reviews Q&A
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      1-6 of 6 Abreviews or Q&A

      Immunocytochemistry/ Immunofluorescence abreview for Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S5) antibody [8A7]

      Average
      Abreviews
      Abreviews
      abreview image
      Application
      Immunocytochemistry/ Immunofluorescence
      Sample
      Human Cell (Osteosarcoma)
      Permeabilization
      Yes - 0.3% tritonX-100
      Specification
      Osteosarcoma
      Blocking step
      Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
      Fixative
      Methanol
      Read More
      The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

      Abcam user community

      Verified customer

      Submitted May 28 2021

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

      Good
      Abreviews
      Abreviews
      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
      Read More
      The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

      Abcam user community

      Verified customer

      Submitted Feb 21 2013

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

      Excellent
      Abreviews
      Abreviews
      abreview image
      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
      Read More
      The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

      Abcam user community

      Verified customer

      Submitted Jan 19 2012

      Immunocytochemistry/ Immunofluorescence abreview for Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S5) antibody [8A7]

      Excellent
      Abreviews
      Abreviews
      abreview image
      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
      Read More
      The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

      Abcam user community

      Verified customer

      Submitted Jun 07 2010

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

      Good
      Abreviews
      Abreviews
      abreview image
      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%
      Read More
      The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

      Abcam user community

      Verified customer

      Submitted Jan 02 2007

      Question

      Please could you tell me if you have a WB image for ab5401 on a human cell lysate?

      Read More

      Abcam community

      Verified customer

      Asked on Mar 17 2004

      Answer

      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 latest batch failed QC tests and a new batch is in production and should be release by the end of April 2004 - Last update 12the March 04. If you need anything further or any help then please let us know.

      Read More

      Abcam Scientific Support

      Answered on Mar 18 2004

      Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
      For licensing inquiries, please contact partnerships@abcam.com

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