Overview

  • Product nameAnti-Cleaved PARP antibody [E51]
    See all Cleaved PARP primary antibodies
  • Description
    Rabbit monoclonal [E51] to Cleaved PARP
  • SpecificityThis antibody is specific for the p25 cleaved form of human PARP.
  • Tested applicationsSuitable for: WB, IHC-Pmore details
    Unsuitable for: ICC/IF
  • Species reactivity
    Reacts with: Mouse, Rat, Human
    Predicted to work with: Chinese hamster
  • Immunogen

    Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) corresponding to Human Cleaved PARP aa 150-250.

  • Positive control
    • Jurkat cells.
  • General notes

    This product is a recombinant rabbit monoclonal antibody.

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated ‘PUR’ on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

    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.

     

    Alternative versions available:

    Anti-Cleaved PARP antibody (HRP) [E51] (ab194217)

Properties

Applications

Our Abpromise guarantee covers the use of ab32064 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 - 1/10000. Predicted molecular weight: 25 kDa.
IHC-P 1/100.
  • Application notesIs unsuitable for ICC/IF.
  • Target

    • FunctionInvolved 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 similaritiesContains 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 localizationNucleus. 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 PARP antibody [E51] images



    • Predicted band size : 25 kDa

      Lane 1: Wild type HAP1 whole cell lysate (20 µg)
      Lane 2: PARP1 knockout  HAP1 whole cell lysate (20 µg)
      Lane 3: HeLa whole cell lysate (20 µg)
      Lane 4: MCF7 whole cell lysate (20 µg)

      Lanes 1 - 4: Merged signal (red and green). Green - ab32064 observed at 30 kDa. Red - loading control, ab8245, observed at 37 kDa.

      ab32064 was shown to specifically react with PARP1 when PARP1 knockout samples were used. Wild-type and PARP1 knockout samples were subjected to SDS-PAGE.  Ab32064 and ab8245 (Mouse anti GAPDH loading control) were incubated overnight at 4°C at 1/10000 dilutions. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) ab216773 and 680CW Goat anti Mouse secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.

    • All lanes : Anti-Cleaved PARP antibody [E51] (ab32064) at 1/1000 dilution

      Lane 1 : HeLa treated with 1uM Staurosporine for 3 hours whole cell lysates
      Lane 2 : Untreated HeLa whole cell lysates
      Lane 3 : NIH/3T3 treated with 1uM Staurosporine for 3 hours whole cell lysates
      Lane 4 : Untreated NIH/3T3 whole cell lysates

      Lysates/proteins at 20 µg per lane.

      Secondary
      Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

      Predicted band size : 25 kDa
      Observed band size : 25 kDa

      Blocking/Dilution buffer 5% NFDM/TBST

      Exposure time :

      Lane 1,2: 1 second
      Lane 3,4: 8 seconds

    • Immunohistochemical staining of paraffin embedded rat colon with purified ab32064 at a working dilution of 1 in 100. The secondary antibody used is a HRP polymer for rabbit IgG. The sample is counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.

    • All lanes : Anti-Cleaved PARP antibody [E51] (ab32064) at 1/1000 dilution

      Lane 1 : RAW264.7 cell lysate
      Lane 2 : NIH/3T3 cell lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      HRP goat anti-rabbit (H+L) at 1/1000 dilution

      Predicted band size : 25 kDa
      Observed band size : 25 kDa

      Blocking buffer: 5% NFDM/TBST

      Dilution buffer: 5% NFDM/TBST

    • All lanes : Anti-Cleaved PARP antibody [E51] (ab32064) at 1/10000 dilution

      Lane 1 : Untreated Jurkat cell lysate
      Lane 2 : Jurkat cell lysate treated with camptothecin

      Lysates/proteins at 10 µg per lane.

      Secondary
      HRP goat anti-rabbit (H+L) at 1/1000 dilution

      Predicted band size : 25 kDa
      Observed band size : 25 kDa

      Blocking buffer: 5% NFDM/TBST

      Dilution buffer: 5% NFDM/TBST

    • Immunohistochemical staining of paraffin embedded human ovarian carcinoma with purified ab32064 at a working dilution of 1 in 100. The secondary antibody used is a HRP polymer for rabbit IgG. The sample is counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.


    • Developed using the ECL technique

      Performed under reducing conditions.

      Predicted band size : 25 kDa
      Observed band size : 25 kDa


      Exposure time : 10 seconds

      Jurkat cells were incubated at 37°C for 24 hours with vehicle control (0 μM) and different concentrations of 15-Acetoxyscirpenol (ab142381). Increased expression of cleaved PARP (ab32064) in Jurkat cells correlates with an increase in 15-Acetoxyscirpenol concentration, as described in literature.

      Whole cell lysates were prepared with RIPA buffer (containing protease inhibitors and sodium orthovanadate), 20μg of each were loaded on the gel and the WB was run under reducing conditions. After transfer the membrane was blocked for an hour using 5% BSA before being incubated with ab32064 at 1/10000 dilution and ab8227 at 1 μg/ml overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP (ab97051) at 1/10000 and visualised using ECL development solution.

    • Anti-Cleaved PARP antibody [E51] (ab32064) at 1/50000 dilution + MCF7 cell lysate at 100 µg

      Secondary
      HRP-conjugated Goat anti-rabbit IgG polyclonal at 1/50000 dilution
      Developed using the ECL technique

      Performed under reducing conditions.

      Predicted band size : 25 kDa
      Observed band size : 25 kDa


      Exposure time : 15 minutes

      This image is courtesy of an anonymous Abreview

      See Abreview

    • All lanes : Anti-Cleaved PARP antibody [E51] (ab32064) at 1/1000000 dilution

      Lane 1 : Jurkat cell lysate. Untreated.
      Lane 2 : Jurkat cell lysate. Treated with Camptothecin.


      Predicted band size : 25 kDa
      Observed band size : 25 kDa
    • All lanes : Anti-Cleaved PARP antibody [E51] (ab32064) at 1/1000 dilution

      Lane 1 : PC-12 treated with 1uM Staurosporine for 3 hours whole cell lysates
      Lane 2 : Untreated PC-12 whole cell lysates

      Lysates/proteins at 20 µg per lane.

      Secondary
      Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

      Predicted band size : 25 kDa
      Observed band size : 25 kDa


      Exposure time : 30 seconds

      Blockinng/Diluting buffer 5% NFDM/TBST

    • Immunohistochemical staining of paraffin embedded human breast carcinoma with unpurified ab32064 at a 1:100 dilution.

    References for Anti-Cleaved PARP antibody [E51] (ab32064)

    This product has been referenced in:
    • Qin Q  et al. miR-134 inhibits non-small cell lung cancer growth by targeting the epidermal growth factor receptor. J Cell Mol Med N/A:N/A (2016). IHC . Read more (PubMed: 27241841) »
    • Song J  et al. Cordyceps militaris induces tumor cell death via the caspase-dependent mitochondrial pathway in HepG2 and MCF-7 cells. Mol Med Rep 13:5132-40 (2016). WB ; Human . Read more (PubMed: 27109250) »

    See all 27 Publications for this product

    Product Wall

    The antigen retrieval was heat-mediated with citrate buffer, pH 6.0. The laboratory uses a pressure cooker but a microwave, steamer, or waterbath should work too.


    The concentration for the aforementioned lot is 0.3770 mg/ml

    Thank you for contacting Abcam regarding ab32064.


    I have confirmed with the laboratory that we currently do not have any data demonstrating the use of this antibody with a fluorescently conjugated secondary antibody. However, we expect ...

    Read More

    Thank you for contacting us. We have several antibodies which can be used to detect PARP in Western blotting. From the study I believe you wish to detect both the cleaved (C-terminal fragment) and the full length PARP form in mouse samples? Thi...

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