Overview

  • Product name

    Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S2) antibody
    See all RNA polymerase II CTD repeat YSPTSPS primary antibodies
  • Description

    Rabbit polyclonal to RNA polymerase II CTD repeat YSPTSPS (phospho S2)
  • Host species

    Rabbit
  • Tested applications

    Suitable for: IHC-P, WB, ELISAmore details
  • Species reactivity

    Reacts with: Mouse, Human
    Predicted to work with: Rat, Rabbit, Cow, Dog, Pig, Chimpanzee, Drosophila melanogaster, Plants, Zebrafish, Rhesus monkey, Gorilla, Orangutan, Xenopus tropicalis
  • Immunogen

    Synthetic peptide corresponding to Human RNA polymerase II CTD repeat

    YSPTSPS

    (C terminal) (phospho S2).
    Sequence:

    YSPTSPS


    Database link: P24928

  • Positive control

    • Whole cell lysate from HeLa cells

Properties

Applications

Our Abpromise guarantee covers the use of ab70324 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
IHC-P 1/200 - 1/1000. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
WB 1/2500 - 1/20000. Detects a band of approximately 270 kDa.
ELISA Use at an assay dependent concentration.

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.
  • Information by UniProt
  • Database links

    see all
  • 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

  • All lanes : Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S2) antibody (ab70324) at 0.02 µg/ml

    Lane 1 : Whole cell lysate from HeLa cells with no peptide
    Lane 2 : Whole cell lysate from HeLa cells with non-phosphorylated peptide
    Lane 3 : Whole cell lysate from HeLa cells with phosphorylated S2 peptide
    Lane 4 : Whole cell lysate from HeLa cells with phosphorylated S5 peptide

    Lysates/proteins at 50 µg per lane.

    Observed band size: 270 kDa
    why is the actual band size different from the predicted?

  • Serially diluted ab70324 was bound to immobilised phospho- or control peptides (1 microgram per mL). The antibody was detected by HRP-labelled goat anti-rabbit IgG (ab97080; diluted 50000 times) and signal was developed by TMB substrate.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human ovarian carcinoma mock phosphatase treated section (left) and calf intestinal phosphatase-treated section (right) tissues labelling RNA polymerase II CTD repeat YSPTSPS (phospho S2) with ab70324 at 1/500 (0.4µg/ml). Detection: DAB.

References

This product has been referenced in:

  • Syn NL  et al. Pan-CDK inhibition augments cisplatin lethality in nasopharyngeal carcinoma cell lines and xenograft models. Signal Transduct Target Ther 3:9 (2018). Read more (PubMed: 29666673) »
See 1 Publication for this product

Customer reviews and Q&As

1-3 of 3 Abreviews or Q&A

Question
Answer

Thank you for contacting Abcam and thank you for your patience while I consulted my coleagues about your enquiry. Below is the response that I received from a colleague: "I would suggest using the non phosphorylated form of Pol2 as a positive control for ChIP. The phosophorylation state may change so detecting the protein would be more appropriate. I suppose it depends on what experiments they are doing, in this case the experimental cells may have higher phosporylation levels than the control creating artifactual results so they should be careful. They may need to do a second control to be sure." I hope this answers your questions and if I can be of anymore help, please let me know.

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Question
Answer

Please accept my apologies for misunderstanding your original email. I have to admit that I am no expert in ChIP assays but I have sent an email to ask my colleagues and I will let you know what they say. I see no reason why you could not use S2 phospho Pol2 as a positive control but as I said I am no expert. As to the discount code, under normal circumstances we have to issue the code before the original purchase but since I have already offered it to you, I will make an exception. The discount code and details of how to use it are below. Once again, please accept my apologies for the original misunderstanding and I will let you know what I hear back from my colleagues. If there is anything else I can do in the mean time, please let me know and I look forward to reading your Abreview. DISCOUNT CODE: ******** Expiration date: ******** I am very pleased to hear you would like to accept our offer and test ab70324 in ChIP. This code will give you 1 free primary antibody before the expiration date. To redeem this offer, please submit an Abreview for ChIP and include this code in the “Additional Comments” section so we know the Abreview is for this promotion. For more information on how to submit an Abreview, please visit the site: www.abcam.com/Abreviews. Remember, we publish both positive and negative Abreviews on our datasheets so please submit the results of your tests. The code will be active once the Abreview has been submitted and can be redeemed in one of the following ways: 1) Call to place your order and mention the code to our customer service department; 2) Include the code in your fax order; 3) Place your order on the web and enter the promotional code. Any feedback that you can provide will be greatly appreciated, whether positive or negative. If you have any further questions, please do not hesitate to contact us. We look forward to receiving your Abreview and wish you luck with your research. The terms and conditions applicable to this offer can be found here: www.abcam.com/collaborationdiscount.

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Answer

Thank you for contacting Abcam about ab70324. As long as there is a sufficient concentration of your sample potein present, the anti-RNA polymerase II CTD repeat YSPTSPS (phospho S2) antibody should work. As it has not been tested in ChIP, we cannot gaurantee that it will work for you, but I can offer you our testing discount. The details of which are below: As ab70324 has not been tested in ChIP. Therefore, I can offer a discount off a future purchase if you buy ab70324 now, test it in ChIP and submit feedback to us in the form of an Abreview. It doesn’t matter whether the Abreview is positive or negative, we would just really like to receive your feedback. The discount would be to the value of 1 free primary antibody. If you are interested in this offer, please follow these steps: 1. Reply to this e-mail to let me know that you would like to proceed and test ab70324 in ChIP. I will then send a discount code. This code must be issued before purchasing ab70324 so please wait for my reply before ordering. 2. Purchase ab70324 either by phone, fax, or online (www.abcam.com). 3. Test it in ChIP. 4. Let us know the results, positive or negative, using our Abreview system (this will take about 10 minutes and images are great if you have them!). To find out how to submit an Abreview, please visit: https://www.abcam.com/abreviews. 5. After the review is submitted to us, the discount code becomes active. Simply place your new order by phone, fax, or on the web and mention the discount code. The discount can be redeemed for any primary antibody ordered and the discount code is valid for 4 months after issue. We are always pleased to obtain feedback about our products and any information is greatly appreciated! Even if ab70324 turns out to be unsuitable for ChIP you will still receive the discount on your next purchase after your Abreview has been submitted. Please let me know if you have any questions about this offer and I would be happy to help you further. The Terms and Conditions of this offer can be found at: www.abcam.com/collaborationdiscount.

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