Overview

  • Product name
    Anti-Cleaved PARP1 antibody
    See all Cleaved PARP1 primary antibodies
  • Description
    Rabbit polyclonal to Cleaved PARP1
  • Host species
    Rabbit
  • Specificity
    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.
  • Tested applications
    Suitable for: WB, ICC/IFmore details
  • Species reactivity
    Reacts with: Human
  • Immunogen

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

Properties

Applications

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.

Application Abreviews Notes
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.

Target

  • Function
    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.
  • Sequence similarities
    Contains 1 BRCT domain.
    Contains 1 PARP alpha-helical domain.
    Contains 1 PARP catalytic domain.
    Contains 2 PARP-type zinc fingers.
  • Post-translational
    modifications
    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.
  • Cellular localization
    Nucleus. Nucleus, nucleolus. Localizes at sites of DNA damage.
  • Information by UniProt
  • Database links
  • Alternative names
    • 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 antibody
    • Poly ADP ribose polymerase 1 antibody
    • Poly[ADP-ribose] synthase 1 antibody
    • PPOL antibody
    • SCA1 antibody
    see all

References

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

Customer reviews and Q&As

1-3 of 3 Abreviews or Q&A

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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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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Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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