Overview

  • Product name

    Anti-Cleaved PARP1 antibody [4B5BD2]
    See all Cleaved PARP1 primary antibodies
  • Description

    Mouse monoclonal [4B5BD2] to Cleaved PARP1
  • Host species

    Mouse
  • 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. This information is considered to be commercially sensitive.

  • 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.

    If you require this antibody in a particular buffer formulation or a particular conjugate for your experiments, please contact orders@abcam.com or you can find further information here.

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
    • 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

Images

  • 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

    Lanes 1 & 3 : untreated HeLa cells
    Lanes 2 & 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:

  • Hou Y  et al. Genetic ablation of TAZ induces HepG2 liver cancer cell apoptosis through activating the CaMKII/MIEF1 signaling pathway. Onco Targets Ther 12:1765-1779 (2019). Read more (PubMed: 30881030) »
  • Huang D  et al. Oxaliplatin and infliximab synergize to induce regression of colon cancer. Oncol Lett 15:1517-1522 (2018). Read more (PubMed: 29434844) »
See all 4 Publications for this product

Customer reviews and Q&As

1-5 of 5 Abreviews or Q&A

Application
Immunocytochemistry/ Immunofluorescence
Sample
Rat Cell (Primary cardiac fibroblasts)
Permeabilization
Yes - 1% triton for 4 minutes
Specification
Primary cardiac fibroblasts
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 4% · Temperature: 25°C
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted Sep 20 2018

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

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

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

Dr. Kirk Mcmanus

Verified customer

Submitted Feb 20 2013

Answer

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 moment it is not mentioned there.

We do however guarantee this antibody ab110315 on human derived cells. Please do let me know in what application and on what species all you need an anti cleaved PARP antibody, and I can see whether we do have a suitable antibody for you. We do have for example the ab32064, which has been tested on mouse tissue. https://www.abcam.com/index.html?datasheet=110315 (or use the following: https://www.abcam.com/index.html?datasheet=110315).

I hope this information is helpful. Please do not hesitate to contact us for further information.

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