Anti-Cleaved PARP1 抗体 (ab2317)

Submit a review Q&A (3)References (11)

製品の概要

  • 製品名

    Anti-Cleaved PARP1 antibody
    Cleaved PARP1 一次抗体 製品一覧

  • 製品の詳細

    Rabbit polyclonal to Cleaved PARP1

  • 由来種

    Rabbit

  • 特異性

    This antibody recognizes only the large fragment of PARP1 (89 kDa) and does not react with full length PARP1. The immunogen does not contain Asp214.

  • アプリケーション

    適用あり: WB, ICC/IFmore details

  • 種交差性

    交差種: Human

  • 免疫原

    Synthetic peptide corresponding to Human Cleaved PARP1 (N terminal). N-terminal residues of the catalytic domain of human PARP1.
    Database link: NP_001609

製品の特性

アプリケーション

Our Abpromise guarantee covers the use of ab2317 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 Abreviews 特記事項
WB Use a concentration of 1 µg/ml. Detects a band of approximately 89 kDa.
ICC/IF Use a concentration of 10 - 20 µg/ml.

ターゲット情報

  • 機能

    Involved in the base excision repair (BER) pathway, by catalyzing the poly(ADP-ribosyl)ation of a limited number of acceptor proteins involved in chromatin architecture and in DNA metabolism. This modification follows DNA damages and appears as an obligatory step in a detection/signaling pathway leading to the reparation of DNA strand breaks. Mediates the poly(ADP-ribosyl)ation of APLF and CHFR. Positively regulates the transcription of MTUS1 and negatively regulates the transcription of MTUS2/TIP150. With EEF1A1 and TXK, forms a complex that acts as a T-helper 1 (Th1) cell-specific transcription factor and binds the promoter of IFN-gamma to directly regulate its transcription, and is thus involved importantly in Th1 cytokine production. Required for PARP9 and DTX3L recruitment to DNA damage sites. PARP1-dependent PARP9-DTX3L-mediated ubiquitination promotes the rapid and specific recruitment of 53BP1/TP53BP1, UIMC1/RAP80, and BRCA1 to DNA damage sites.

  • 配列類似性

    Contains 1 BRCT domain.
    Contains 1 PARP alpha-helical domain.
    Contains 1 PARP catalytic domain.
    Contains 2 PARP-type zinc fingers.

  • 翻訳後修飾

    Phosphorylated by PRKDC and TXK.
    Poly-ADP-ribosylated by PARP2. Poly-ADP-ribosylation mediates the recruitment of CHD1L to DNA damage sites.
    S-nitrosylated, leading to inhibit transcription regulation activity.

  • 細胞内局在

    Nucleus. Nucleus, nucleolus. Localizes at sites of DNA damage.
  • Information by UniProt

  • 参照データベース

    • 別名

      • ADP ribosyltransferase diphtheria toxin like 1 antibody
      • ADP ribosyltransferase NAD(+) antibody
      • ADP-ribosyltransferase diphtheria toxin-like 1 antibody
      • ADPRT 1 antibody
      • ADPRT antibody
      • ADPRT1 antibody
      • APOPAIN antibody
      • ARTD1 antibody
      • NAD(+) ADP-ribosyltransferase 1 antibody
      • PARP antibody
      • PARP-1 antibody
      • PARP1 antibody
      • PARP1_HUMAN antibody
      • Poly [ADP-ribose] polymerase 1 (PARP-1) antibody
      • Poly [ADP-ribose] polymerase 1 antibody
      • Poly ADP ribose polymerase 1 antibody
      • Poly(ADP ribose) polymerase antibody
      • Poly[ADP-ribose] synthase 1 antibody
      • PPOL antibody
      • SCA1 antibody
      see all

    参考文献

    This product has been referenced in:

    • Gorelick-Ashkenazi A  et al. Caspases maintain tissue integrity by an apoptosis-independent inhibition of cell migration and invasion. Nat Commun 9:2806 (2018). Read more (PubMed: 30022065) »
    • Williams AA  et al. Apoptotic Activity of MeCP2 Is Enhanced by C-Terminal Truncating Mutations. PLoS One 11:e0159632 (2016). Read more (PubMed: 27442528) »
    See all 11 Publications for this product

    ab2317 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。

    レビューと Q&A

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    1-3 of 3 Abreviews or Q&A

    Question

    I am sorry it has taken me a while to respond to your last email regarding the ab2317 rabbit-anti-active PARP antibody, but I wanted to test out your suggestions before getting back to you.

    I tried the different fixation and blocking methods that you suggested in your previous email but have still had no luck with seeing immunoreactivity of the antibody. Would it be possible to have another batch of antibody sent to us for me to test out?

    Thank you again for your time and help. I look forward to hearing from you soon.

    Read More
    Answer

    Thank you for your reply.

    I am sorry this product did not perform as stated on the datasheet and for the inconvenience this has caused. As requested, I have issued a free of charge replacement.

    To check the status of the order please contact our Customer Service team and reference this number.

    Please note that this free of charge replacement vial is also covered by our Abpromise guarantee. Should you still be experiencing difficulties, or if you have any further questions, please do not hesitate to let us know.

    I wish you the best of luck with your research.

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    Question

    Thank you so much for your prompt reply. In response to your question, I am staining dissected pupal epidermal samples in order to image dendritic arborization neurons using confocal microscopy. The ab2317 antibody has been successfully used before in other labs for this very purpose, and has been documented in several publications. Below is a detailed protocol that I have been following for the sample preparation and protocol.


    Staining Procedure:

    7. Immediately fix samples in 4% PFA diluted in 1X PBS for 30 min at room temperature (I have increased the fixation time to at most 1 hr during troubleshooting).

    8. Rinse fixed pupae 3x, 5 minutes each in 1X PBT-X (1X PBS + 0.3% Triton-X-100).

    9.Block samples in 1X PBT-X + 5% normal goat serum for 1 hr at 4 degrees Celsius.

    10. Incubate with 10 ug/mL rabbit-anti-cleaved PARP diluted in 5% normal goat serum/PBT-X overnight at 4 degrees Celsius.

    11. The next day, remove antibody dilution and wash samples 3x, 10 minutes each in 1X PBT-X at room temperature.

    12. Incubate in secondary antibody 1:200 goat-anti-rabbit-Alexa 488 diluted in 5% normal goat serum/PBT-X for 2 hrs at room temperature.(also have tried Alexa 568 and Cy5-conjugated secondary antibodies)

    13. Wash samples 3X, 10 minutes each in 1X PBT-X.

    14. Equilibrate samples in glycerol-based mounting media for at least 30 minutes.

    15. Separate epidermis from pupal case and mount on cover slip and slide. Store samples at -20 degrees.

    I am also co-staining these samples with a mouse-anti-GFP antibody-this staining has been constantly strong and clean which suggests that my sample preparation and staining procedure is working. I'd appreciate any help on this matter. I have been troubleshooting this staining for months and it would be great to have some advice. One other thing I was wondering is if anyone has had any issues with the particular lot of antibody that I purchased or with this antibody in general.

    Thank you again for getting back to me so quickly and for your help. I look forward to hearing from you soon.

    Read More
    Answer

    Thank you for your reply with the protocol information.


    Based on the protocol information you provided, it seems that you are using relatively standard methods that should be compatible with the antibody. One possibility would be to try a different fixative, such as methanol or acetone. Additionally, blocking (and diluting your antibodies)with 3% BSA rather than serum may also help your staining.


    I have checked our records and we have not had other users report any difficulties using this antibody. I am happy to offer a replacement or credit if these suggestions do not improve your results. I look forward to your reply so that I may assist you further.

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    Question

    We purchased an antibody froagainst cleaved PARP (It is the rabbit polyclonal to cleaved PARP ab2317). However, after many attempts for immunofluorescence detection, I have been unable to see any immunoreactivity of this antibody using the concentrations recommended in the data sheet supplied with the antibody (dilution of 1:10-1:20, both dilutions were attempted) and with conditions successfully used in previously published reports. I was wondering if anybody has reported issues with this antibody recently. I am using standard staining techniques and using it in Drosophila pupae expressing a UAS-CD8::PARP-Venus reporter driven by Gal4 expression, as similarly used in Williams et al 2006, Nature Neuroscience and Rumpf et al 2011, Development. Any suggestions/information would be much appreciated. Thank you in advance and I look forward to hearing from you.

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    Answer

    Thank you for contacting Abcam regarding ab2317.


    I am sorry that you have been experiencing difficulties with this antibody in your experiments. I was hoping you would provide me with some additional information so that I may assist you in resolving this issue. In particular, are you staining whole pupae, tissue sections, or cells in culture? As the antibody is specifically validated for ICC I want to be sure you are using this application. Would you also send me a detailed summary of your protocol and sample preparation? If possible, I would be happy to offer some protocol recommendations, or alternatively a replacement or credit per our Abpromise guarantee.


    I look forward to your reply so that I may help you further. Please do not hesitate to contact us if you have any additional questions.

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