Validated using a knockout cell line
Recombinant
RabMAb

Anti-Cleaved PARP1 antibody [Y34] (ab32561)

Overview

  • Product name
    Anti-Cleaved PARP1 antibody [Y34]
    See all Cleaved PARP1 primary antibodies
  • Description
    Rabbit monoclonal [Y34] to Cleaved PARP1
  • Specificity
    This antibody is specific for p85 cleaved form of PARP1.
  • Tested applications
    Suitable for: WB, Flow Cyt, IP, ICC/IFmore details
    Unsuitable for: IHC
  • Species reactivity
    Reacts with: Human
  • Immunogen

    Synthetic peptide within Human Cleaved PARP1 aa 200-300. The exact sequence is proprietary. Residues following the cleavage of site.

  • Positive control
    • Jurkat cell lysate.
  • General notes

    This product is a recombinant rabbit monoclonal antibody.

    Produced using Abcam’s RabMAb® technology. RabMAb® technology is covered by the following U.S. Patents, No. 5,675,063 and/or 7,429,487.

    A trial size is available to purchase for this antibody.

    Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.

Applications

Our Abpromise guarantee covers the use of ab32561 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 1/1000. Predicted molecular weight: 85 kDa.
Flow Cyt 1/50. ab172730-Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
IP 1/50.
ICC/IF 1/500.
  • Application notes
    Is unsuitable for IHC.
  • 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

    Anti-Cleaved PARP1 antibody [Y34] images

    • Primary ab 1/50 dilution (0.5μg / Red). Secondary ab Goat anti rabbit IgG (FITC). Secondary ab concentration 1/150 dilution. Cell line Jurkat (human acute T cell leukemia) treated with (Right) or without (Left) 4μM Camptothecin for 5h. Fixative 4% paraformaldehyde. Datasheet comment Flow cytometric analysis of apoptotic and non-apoptotic Jurkat cells using anti-cleaved PARP1 RabMAb (ab32561). Jurkat cells were either left untreated (A) or treated with camptothecin (4 uM, 5 hr) to induce apoptosis (B). Cells were fixed and permeabilized, and then stained with anti-cleaved PARP1. The results indicate that 43% of cells were positive for cleaved PARP1 (B, M2) after treatment, compared to 9% positive without treatment (A, M2).



    • Predicted band size : 85 kDa

      Lane 1: Wild type HAP1 (untreated) whole cell lysate (20 µg)
      Lane 2: PARP1 (untreated) knockout HAP1 (untreated) whole cell lysate (20 µg)
      Lane 3: HeLa (untreated) whole cell lysate (20 µg)
      Lane 4: HAP1 (staurosporin treated, 1 u M, 4 hr) whole cell lysate (20 µg)
      Lane 5: PARP1 (staurosporin treated, 1 uM, 4 hr) knockout HAP1 whole cell lysate (20 µg)
      Lane 6: HeLa (staurosporin treated, 1 uM, 4 hr) whole cell lysate (20 µg)

      Lanes 1 - 6: Merged signal (red and green). Green - ab32561 observed at 100 kDa. Red - loading control, ab8245, observed at 37 kDa.

      ab32561 was shown to specifically react with PARP1 (untreated) when PARP1 (untreated) knockout samples were used. Wild-type and PARP1 (untreated) knockout samples were subjected to SDS-PAGE. Ab32561 and ab8245 (Mouse anti GAPDH loading control) were incubated overnight at 4°C at 1000 dilution and 1/10000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.              

    • ab32561 staining Cleaved PARP1 in HeLa cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with formaldehyde and permeabilized with 0.5% Triton X-100 in PBS. Samples were incubated with primary antibody (1/500 in PBS) for 1 hour at 22°C. ab150081, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG polyclonal (1/200) was used as the secondary antibody. Counterstained with DAPI.

      See Abreview

    • All lanes : Anti-Cleaved PARP1 antibody [Y34] (ab32561) at 1/1000 dilution

      Lane 1 : Un-treated Jurkat cell lysate.
      Lane 2 : Jurkat cell lysate treated with Camptothecin.


      Predicted band size : 85 kDa
      Observed band size : 85 kDa
    • Immunohistochemical analysis of OVCAR-3 tumour xenografts in nude mice, staining cleaved PARP1 with ab32561.

      Antigen retrieval was performed via heat mediation in a citrate buffer. Samples were incubated with primary antibody (1/100) overnight at 4°C. A biotinyated anti-rabbit IgG (1/150) was used as the secondary antibody and staining was detected using DAB.

    References for Anti-Cleaved PARP1 antibody [Y34] (ab32561)

    This product has been referenced in:
    • Jiao S  et al. Inhibition of CYFIP2 promotes gastric cancer cell proliferation and chemoresistance to 5-fluorouracil through activation of the Akt signaling pathway. Oncol Lett 13:2133-2140 (2017). WB ; Human . Read more (PubMed: 28454373) »
    • Peh J  et al. The Combination of Vemurafenib and Procaspase-3 Activation Is Synergistic in Mutant BRAF Melanomas. Mol Cancer Ther 15:1859-69 (2016). Read more (PubMed: 27297867) »

    See all 14 Publications for this product

    Product Wall

    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Human Cell (HeLa)
    Permeabilization
    Yes - 0.5% Triton-X100 in PBS
    Specification
    HeLa
    Fixative
    Formaldehyde
    Username

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    Verified customer

    Submitted Nov 05 2014

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

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