Anti-Cleaved PARP1 antibody [4B5BD2] (ab110315)

Overview

  • Product name
    Anti-Cleaved PARP1 antibody [4B5BD2]
    See all Cleaved PARP1 primary antibodies
  • Description
    Mouse monoclonal [4B5BD2] to Cleaved PARP1
  • Specificity
    ab110315 reacts with the N-terminal end formed by the cleavage adjacent to Asp214; it thus recognizes the apoptosis-specific 89 kDa catalytic domain fragment, but it does not recognize the full-length PARP1 or the 24 kDa DNA binding domain fragment.
  • Tested applications
    Suitable for: WB, ICC/IF, In-Cell ELISA, Flow Cytmore details
  • Species reactivity
    Reacts with: Human
  • Immunogen

    Synthetic peptide corresponding to Human Cleaved PARP1 (N terminal).
    Sequence:

    GVDEVAKKKSKKEK-C

  • Positive control
    • Staurosporine-treated HeLa and HL60 cells
  • General notes

    This antibody clone is manufactured by Abcam.

    This monoclonal antibody to cleaved PARP1 has been knockout validated in Western blot. The expected band for cleaved PARP1 was observed in wild type cells and the band was not seen in knockout cells.

    Product was previously marketed under the MitoSciences sub-brand.

Properties

Applications

Our Abpromise guarantee covers the use of ab110315 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 0.25 - 1 µg/ml. Predicted molecular weight: 113 kDa.
ICC/IF Use a concentration of 1 µg/ml.
In-Cell ELISA Use a concentration of 1 µg/ml.
Flow Cyt Use a concentration of 1 µg/ml.

ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.

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

Images



  • Predicted band size : 113 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 (staurosporine treated, 1 uM, 4 hr) whole cell lysate (20 µg)
    Lane 5: PARP1 (staurosporine treated, 1 uM, 4 hr) knockout HAP1 whole cell lysate (20 µg)
    Lane 6: HeLa (staurosporine treated, 1 uM, 4 hr) whole cell lysate (20 µg)

    Lanes 1 - 6: Merged signal (red and green). Green - ab110315 observed at 100 kDa. Red - loading control, ab181602, observed at 37 kDa

    ab110315 detected the expected band for cleaved PARP1 in wild type HAP1 cells treated with staurosporine and the band was not seen in PARP1 knockout cells treated with staurosporine. Wild-type and PARP1 knockout samples were subjected to SDS-PAGE. ab110315 and ab181602 (Rabbit anti GAPDH loading control) were incubated overnight at 4°C at 1 ug/ml and 1/10000 dilution respectively. Blots were developed with Goat anti-Mouse IgG H&L (IRDye® 800CW) preabsorbed ab216772 and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preabsorbed ab216777 secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.

  • Immunocytochemistry images of stained untreated (A) and 4 hours 1 µM Staurosporine-treated (B) Human HeLa cells. The cells were paraformaldehyde fixed (4%, 20 minutes) and Triton X-100 permeabilized (0.1%, 15 minutes). The cells were incubated with 1.0 µg/ml ab110315 for 2 hours at room temperature or over night at 4°C. 10% goat serum was used as the blocking agent for all blocking steps. The secondary antibody was Alexa Fluor® 488 goat anti-mouse IgG (H+L) (in green) used at 2.0 µg/ml for 2 hours. DAPI was used to stain the cell nuclei (in red). Heat induced antigen retrieval (0.1 M Tris-HCl, 5% urea, pH 9.5 for 5 min at 95°C) improves signal. Note that the ab110315 labels only condensed and/or fragmented nuclei of apoptotic Staurosporine-treated cells.

  • Lanes 1 - 2 : Antibody that recognizes full-length PARP1
    Lanes 3 - 4 : Anti-Cleaved PARP1 antibody [4B5BD2] (ab110315) at 1 µg/ml

    Lane 1 : untreated HeLa cells
    Lane 2 : HeLa cells treated with 1 µM Staurosporinefor 4 hours
    Lane 3 : untreated HeLa cells
    Lane 4 : HeLa cells treated with 1 µM Staurosporinefor 4 hours

    Lysates/proteins at 20 µg per lane.


    Predicted band size : 113 kDa

    Western Blot analysis using ab110315 antibody and 20 µg of untreated (CON) or 4 hours 1 µM Staurosporine-treated (STS) HeLa cells. Blots were incubated with an antibody that recognizes both the full-length PARP1 and its 89 kDa fragment (left panel), or 1.0 µg/mL PARP1 (cleaved) antibody (ab110315) (right panel). Appropriate HRP-conjugated secondary antibodies followed by ECL detection were used. Note that the MS777 antibody recognizes the apoptosis-specific 89 kDa fragment of PARP1 but it does not recognize the full-length PARP1.

  • In-Cell ELISA (ICE) using ab110315 on HeLa cells treated with Staurosporine to induce apoptosis. HeLa cells were seeded overnight (50,000 cells/well), treated for 4 hours with 1 µM Staurosporine or with the drug vehicle (DMSO), fixed for Detaching Adherent Cells and analyzed.
  • Flow cytometry analysis of apoptosis using ab110315. HL-60 cells were treated with 1 µM Staurosporin for 4 hours (blue) or vehicle control (red). Control cells were also stained with an equal amount of an isotype control antibody (black).

References

This product has been referenced in:
  • Arosh JA  et al. Molecular and preclinical basis to inhibit PGE2 receptors EP2 and EP4 as a novel nonsteroidal therapy for endometriosis. Proc Natl Acad Sci U S A 112:9716-21 (2015). IHC-Fr ; Mouse . Read more (PubMed: 26199416) »

See 1 Publication for this product

Customer reviews and Q&As

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - whole cell (epithelial ovacar cell line)
Gel Running Conditions
Reduced Denaturing (7.5%)
Loading amount
100 µg
Treatment
0-200uM Perifosine
Specification
epithelial ovacar cell line
Blocking step
Milk as blocking agent for 12 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C
Username

Abcam user community

Verified customer

Submitted Jul 26 2016

Application
Western blot
Sample
Rat Cell lysate - other (Gonad)
Gel Running Conditions
Reduced Denaturing (10)
Loading amount
75 µg
Specification
Gonad
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Abcam user community

Verified customer

Submitted Sep 12 2013

Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (HeLa)
Permeabilization
Yes - 0.5% Triton X-100 in PBS
Specification
HeLa
Fixative
Paraformaldehyde
Username

Dr. Kirk Mcmanus

Verified customer

Submitted Feb 20 2013

Thank you very much for your inquiry.

I am sorry to confirm, that the laboratory has tested this antibody on mouse tissue and has obtained negative results. We will try to add this information to our datasheets - I apologize that for the mome...

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