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  1. Link

    human-rna-polymerase-ii-ctd-repeat-ysptsps-phospho-s5-peptide-ab18488.pdf

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Epigenetics and Nuclear Signaling Transcription Polymerase associated factors Pol II Transcription
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Human RNA polymerase II CTD repeat YSPTSPS (phospho S5) peptide (ab18488)

  • Datasheet
Reviews (4)Q&A (1)References (4)

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Abpromise

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Dot Blot - Human RNA polymerase II CTD repeat YSPTSPS (phospho S5) peptide (ab18488)

    Key features and details

    • Suitable for: WB, Blocking

    製品の詳細

    • 製品名

      Human RNA polymerase II CTD repeat YSPTSPS (phospho S5) peptide
      RNA polymerase II CTD repeat YSPTSPS タンパク質・ペプチド 製品一覧
    • アクセッション番号

      P24928
    • Animal free

      No
    • 由来

      Synthetic
      • 生物種

        Human
      • 修飾

        phospho S5

    関連製品

    • Corresponding Antibody

      • Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S5) antibody (ab5131)

    特性

    Our Abpromise guarantee covers the use of ab18488 in the following tested applications.

    The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

    • アプリケーション

      Western blot

      Blocking - Blocking peptide for Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S5) antibody (ab5131)

    • 製品の状態

      Liquid
    • 備考

      This peptide is 2 repeats of the YSPTSPS repeated motif found at the C-terminal of RNA polymerase II.

      - First try to dissolve a small amount of peptide in either water or buffer. The more charged residues on a peptide, the more soluble it is in aqueous solutions.
      - If the peptide doesn’t dissolve try an organic solvent e.g. DMSO, then dilute using water or buffer.
      - Consider that any solvent used must be compatible with your assay. If a peptide does not dissolve and you need to recover it, lyophilise to remove the solvent.
      - Gentle warming and sonication can effectively aid peptide solubilisation. If the solution is cloudy or has gelled the peptide may be in suspension rather than solubilised.
      - Peptides containing cysteine are easily oxidised, so should be prepared in solution just prior to use.

    • Concentration information loading...

    前処理および保存

    • 保存方法および安定性

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

      Information available upon request.

    関連情報

    • 別名

      • DNA directed RNA polymerase II A
      • DNA-directed RNA polymerase II largest subunit RNA polymerase II 220 kd subunit
      • DNA-directed RNA polymerase II subunit A
      • DNA-directed RNA polymerase II subunit RPB1
      • DNA-directed RNA polymerase III largest subunit
      • hRPB220
      • hsRPB1
      • POLR2
      • Polr2a
      • POLRA
      • Polymerase (RNA) II (DNA directed) polypeptide A
      • Polymerase (RNA) II (DNA directed) polypeptide A 220kDa
      • RNA polymerase II subunit B1
      • RNA-directed RNA polymerase II subunit RPB1
      • RPB1
      • RPB1_HUMAN
      • RPBh1
      • RpIILS
      • RPO2
      • RPOL2
      see all
    • 機能

      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.
    • 配列類似性

      Belongs to the RNA polymerase beta' chain family.
    • ドメイン

      The C-terminal domain (CTD) serves as a platform for assembly of factors that regulate transcription initiation, elongation, termination and mRNA processing.
    • 翻訳後修飾

      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.
    • 細胞内局在

      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

    画像

    • Dot Blot - Human RNA polymerase II CTD repeat YSPTSPS (phospho S5) peptide (ab18488)
      Dot Blot - Human RNA polymerase II CTD repeat YSPTSPS (phospho S5) peptide (ab18488)This image is courtesy of an anonymous Abreview
      Dot blot analysis of ab18488 - RNA polymerase II CTD repeat YSPTSPS peptide - phospho S5.

      0.1 µg peptide was spotted onto the membrane and blocked with 2% BSA for 1 hour at room temperature, before detection with ab5131 at 1/3000 dilution.

      See Abreview

    プロトコール

    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

    データシートおよび資料

    • Datasheet
  • 参考文献 (4)

    ab18488 を使用した論文を発表された方は、こちらまでお知らせください。データシートに掲載させていただきます。

    ab18488 は 4 報の論文で使用されています。

    • Berletch JB  et al. Escape from X inactivation varies in mouse tissues. PLoS Genet 11:e1005079 (2015). PubMed: 25785854
    • Chan EA  et al. Peripheral subnuclear positioning suppresses Tcrb recombination and segregates Tcrb alleles from RAG2. Proc Natl Acad Sci U S A 110:E4628-37 (2013). PubMed: 24218622
    • Freter R  et al. Adult stem cells exhibit global suppression of RNA polymerase II serine-2 phosphorylation. Stem Cells 28:1571-80 (2010). Blocking . PubMed: 20641035
    • Racanelli AC  et al. A mouse gene that coordinates epigenetic controls and transcriptional interference to achieve tissue-specific expression. Mol Cell Biol 28:836-48 (2008). Blocking . PubMed: 17998333

    レビューと Q&A

    Show All レビュー Q&A
    レビューを送る 質問を送る

    1-5 of 5 Abreviews or Q&A

    Other (In vitro binding assay) Abreview for RNA polymerase II CTD repeat YSPTSPS peptide - phospho S5

    Good
    Abreviews
    Abreviews
    abreview image
    Application
    Other

    Review text: This peptide was mixed with GST tagged recombinant proteins in vitro in binding buffer for 1 hour at room temp. Glutathione coated magnetic beads were then mixed into the buffer and removed after another hour incubation. The GST tagged recombinant proteins and any bound peptide was eluted by incubating with free glutathione. Eluted proteins were analyzed by western blot and peptides analyzed by dotblot.

    This experiment was conducted with ab12793, ab12795 and an unrelated non-Abcam peptide as a negative control.
    Sample: Human Recombinant protein

    Primary antibody (in addition to 'RNA polymerase II CTD repeat YSPTSPS peptide - phospho S5')
    Primary antibody: Abcam primary antibody: Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S5) antibody - ChIP Grade (ab5131)
    Dilution: 1/3000

    Secondary antibody
    Name: Non-Abcam antibody was used: anti-rabbit
    Host species: Goat
    Clonality: Polyclonal
    Conjugation: Horse Radish Peroxidase
    Dilution: 1/20000

    James Thorne

    Verified customer

    投稿 Aug 13 2012

    Other (In vitro pulldown assay) Abreview for RNA polymerase II CTD repeat YSPTSPS peptide - phospho S5

    Good
    Abreviews
    Abreviews
    Application
    Other

    Review text: This peptide was used in an in vitro pulldown assay. It was biotinylated and linked to streptavidin coated magnetic beads. This complex was then mixed with our proteins of interest under a variety of conditions. Proteins which bound to the CTD-S5p peptide were eluted and assessed by western blot.

    This peptide was used along side ab12793, ab12795, and an unrelated non-abcam peptide and binding to each CTD was compared.
    Sample: Human Recombinant protein

    Primary antibody (in addition to 'RNA polymerase II CTD repeat YSPTSPS peptide - phospho S5')
    Primary antibody: None used

    Secondary antibody
    Secondary antibody: None used

    Abcam user community

    Verified customer

    投稿 Aug 07 2012

    Western blot Abreview for RNA polymerase II CTD repeat YSPTSPS peptide - phospho S5

    Good
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot

    Review text: We used this peptide as a positive control for ab5131 (anti S5p-CTD)
    Sample: Human Recombinant protein (ab18488)
    Loading amount: 0.25 µg
    Specification: ab18488
    Gel Running Conditions: Reduced Denaturing (16% tris-tricine (Schagger et al 2006))
    Blocking step: BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 2% · Temperature: 23°C

    Other product details
    Incubation time: 1 hour(s) and 0 minute(s) · Temperature: 23°C

    Primary antibody (in addition to 'RNA polymerase II CTD repeat YSPTSPS peptide - phospho S5')
    Primary antibody: Abcam primary antibody: Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S5) antibody - ChIP Grade (ab5131)

    Secondary antibody
    Name: Non-Abcam antibody was used: anti-rabbit
    Host species: Goat
    Clonality: Polyclonal
    Conjugation: Horse Radish Peroxidase
    Dilution: 1/20000

    Detection
    Detection method: west pico
    Exposure: 30 second(s)
    Bands: Specific: 1.4 kDa
    Positive control: recombinant protein

    Additional data
    Abcam response: Thank you for submitting your Abreview. While you have been able to detect the 1.4 kDa band, we would not typically recommend this peptide for use as a WB positive control because it is very difficult to successfully run and transfer peptides of this size. The peptide is commonly used for blocking purposes.

    Abcam user community

    Verified customer

    投稿 Aug 01 2012

    Other (Dot Blot) Abreview for RNA polymerase II CTD repeat YSPTSPS peptide - phospho S5

    Good
    Abreviews
    Abreviews
    abreview image
    Application
    Other

    Review text: This peptide was used for dot blots to quality control anti-CTD antibodies. It was detected by ab5131 but not with anti-S2-CTD antibody.
    Sample: Human Recombinant protein

    Primary antibody (in addition to 'RNA polymerase II CTD repeat YSPTSPS peptide - phospho S5')
    Dilution: 1/3000

    Secondary antibody
    Name: Non-Abcam antibody was used: anti-rabbit
    Host species: Goat
    Clonality: Polyclonal
    Conjugation: Horse Radish Peroxidase
    Dilution: 1/20000

    Additional data
    Notes: membranes were dotted with 0.1ug of peptide and blocked with 2% bsa at r/t for 1hr prior to primary antibody incubation

    Abcam user community

    Verified customer

    投稿 May 25 2012

    Question

    I would like to know the information of ab18488, ab12795 and ab12793 (peptides) which I received recently. These are the synthetic peptides, so I would like to know the purity and the chemical form of them. Please provide the detail information of them.

    Read More

    Abcam community

    Verified customer

    Asked on Dec 06 2005

    Answer

    Thank you for your enquiry. ab18488 was synthesised as >90 purity by HPLC analysis, although by weight they will contain salts and buffer components. They are supplied as a solution in the buffer stated on the datasheet. ab18488 was synthesised as an amide. ab12795 and ab12793 are both >80% purity and synthesised as carboxyls. I hope this information helps, please do not hesitate to contact us if you need any more advice or information.

    Read More

    Abcam Scientific Support

    Answered on Dec 07 2005

    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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