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Cell Biology Apoptosis Nucleus PARP
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Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-Cleaved PARP1 antibody [Y34] (ab32561)

  • Datasheet
  • SDS
Reviews (1) Submit a question References (60)

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Western blot - Anti-Cleaved PARP1 antibody [Y34] (ab32561)
  • Flow Cytometry (Intracellular) - Anti-Cleaved PARP1 antibody [Y34] (ab32561)
  • Immunoprecipitation - Anti-Cleaved PARP1 antibody [Y34] (ab32561)
  • Western blot - Anti-Cleaved PARP1 antibody [Y34] (ab32561)
  • Immunocytochemistry/ Immunofluorescence - Anti-Cleaved PARP1 antibody [Y34] (ab32561)
  • Anti-Cleaved PARP1 antibody [Y34] (ab32561)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [Y34] to Cleaved PARP1
  • Suitable for: Flow Cyt (Intra), WB, ICC/IF, IP
  • Knockout validated
  • Reacts with: Human

Conjugates logo Related conjugates and formulations

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Overview

  • Product name

    Anti-Cleaved PARP1 antibody [Y34]
    See all Cleaved PARP1 primary antibodies
  • Description

    Rabbit monoclonal [Y34] to Cleaved PARP1
  • Host species

    Rabbit
  • Specificity

    This antibody is specific for p85 cleaved form of PARP1.
  • Tested applications

    Suitable for: Flow Cyt (Intra), WB, ICC/IF, IPmore details
  • 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 whole cell lysate (ab7899). IP: HeLa cell lysate. ICC/IF: HeLa cells
  • General notes

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

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

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
  • Storage buffer

    pH: 7.20
    Preservative: 0.01% Sodium azide
    Constituents: 49% PBS, 50% Glycerol (glycerin, glycerine), 0.05% BSA
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    Y34
  • Isotype

    IgG
  • Research areas

    • Cell Biology
    • Apoptosis
    • Nucleus
    • PARP
    • Epigenetics and Nuclear Signaling
    • Chromatin Modifying Enzymes
    • ADP-ribosylation
    • Epigenetics and Nuclear Signaling
    • DNA / RNA
    • DNA Damage & Repair
    • DNA Damage Response
    • DNA Damage Recognition
    • Epigenetics and Nuclear Signaling
    • DNA / RNA
    • DNA Damage & Repair
    • DNA Damage Response
    • Other
    • Epigenetics and Nuclear Signaling
    • Cell cycle
    • Apoptosis
    • Nuclear
    • Epigenetics and Nuclear Signaling
    • Chromatin Binding Proteins
    • DNA / RNA binding
    • Metabolism
    • Pathways and Processes
    • Metabolism processes
    • Apoptosis
    • Cancer
    • Cell Death
    • Apoptosis
    • Nucleus
    • PARP
    • Cancer
    • Cell Death
    • Apoptosis
    • Metabolism

Associated products

  • Alternative Versions

    • Anti-Cleaved PARP1 antibody [Y34] - BSA and Azide free (ab219953)
    • Alexa Fluor® 488 Anti-Cleaved PARP1 antibody [Y34] (ab237432)
    • Alexa Fluor® 647 Anti-Cleaved PARP1 antibody [Y34] (ab237433)
    • HRP Anti-Cleaved PARP1 antibody [Y34] (ab237434)
  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
    • Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • Positive Controls

    • Jurkat whole cell lysate (ab7899)

Applications

The Abpromise guarantee

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
Flow Cyt (Intra)
1/50.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

WB
1/1000. Predicted molecular weight: 85 kDa.
ICC/IF (1)
Use at an assay dependent concentration.
IP
1/50.
Notes
Flow Cyt (Intra)
1/50.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

WB
1/1000. Predicted molecular weight: 85 kDa.
ICC/IF
Use at an assay dependent concentration.
IP
1/50.

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.
  • Target information above from: UniProt accession P09874 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 142 Human
    • Omim: 173870 Human
    • SwissProt: P09874 Human
    • Unigene: 177766 Human
    • 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

    • Western blot - Anti-Cleaved PARP1 antibody [Y34] (ab32561)
      Western blot - Anti-Cleaved PARP1 antibody [Y34] (ab32561)

      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.              

    • Flow Cytometry (Intracellular) - Anti-Cleaved PARP1 antibody [Y34] (ab32561)
      Flow Cytometry (Intracellular) - Anti-Cleaved PARP1 antibody [Y34] (ab32561)

      Primary ab 1/50 dilution (0.5µg / Red). Secondary abGoat 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 Intracellular 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). 

       

       

    • Immunoprecipitation - Anti-Cleaved PARP1 antibody [Y34] (ab32561)
      Immunoprecipitation - Anti-Cleaved PARP1 antibody [Y34] (ab32561)

      Purified ab32561 at 1/50 dilution (2µg) immunoprecipitating Cleaved PARP1 in HeLa whole cell lysate.
      Lane 1 (input): HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate 10µg
      Lane 2 (+): ab32561 + HeLa whole cell lysate.
      Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab32561 in HeLa whole cell lysate.
      VeriBlot for IP Detection Reagent (HRP) (ab131366) (1/1000 dilution) was used for Western blotting.
      Blocking Buffer and concentration: 5% NFDM/TBST.
      Diluting buffer and concentration: 5% NFDM/TBST.
      Observed band size: 85 kDa

    • Western blot - Anti-Cleaved PARP1 antibody [Y34] (ab32561)
      Western blot - Anti-Cleaved PARP1 antibody [Y34] (ab32561)
      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

    • Immunocytochemistry/ Immunofluorescence - Anti-Cleaved PARP1 antibody [Y34] (ab32561)
      Immunocytochemistry/ Immunofluorescence - Anti-Cleaved PARP1 antibody [Y34] (ab32561)This image is courtesy of an anonymous Abreview

      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

    • Anti-Cleaved PARP1 antibody [Y34] (ab32561)
      Anti-Cleaved PARP1 antibody [Y34] (ab32561)

    Protocols

    To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

    Click here to view the general protocols

    Datasheets and documents

    • SDS download

    • Datasheet download

      Download

    References (60)

    Publishing research using ab32561? Please let us know so that we can cite the reference in this datasheet.

    ab32561 has been referenced in 60 publications.

    • Shi C  et al. lncRNA SNHG14 Plays a Role in Sepsis-Induced Acute Kidney Injury by Regulating miR-93. Mediators Inflamm 2021:5318369 (2021). PubMed: 33505213
    • Wang L  et al. miR-4478 sensitizes ovarian cancer cells to irradiation by inhibiting Fus and attenuating autophagy. Mol Ther Nucleic Acids 23:1110-1119 (2021). PubMed: 33664992
    • Liebl MC  et al. DAZAP2 acts as specifier of the p53 response to DNA damage. Nucleic Acids Res 49:2759-2776 (2021). PubMed: 33591310
    • Ma YS  et al. MicroRNA-499 serves as a sensitizer for lung cancer cells to radiotherapy by inhibition of CK2a-mediated phosphorylation of p65. Mol Ther Oncolytics 21:171-182 (2021). PubMed: 33997273
    • Li X  et al. Circ0120816 acts as an oncogene of esophageal squamous cell carcinoma by inhibiting miR-1305 and releasing TXNRD1. Cancer Cell Int 20:526 (2020). PubMed: 33292234
    View all Publications for this product

    Customer reviews and Q&As

    Show All Reviews Q&A
    Submit a review Submit a question

    Immunocytochemistry/ Immunofluorescence abreview for Anti-Cleaved PARP1 antibody [Y34]

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Human Cell (HeLa)
    Permeabilization
    Yes - 0.5% Triton-X100 in PBS
    Specification
    HeLa
    Fixative
    Formaldehyde
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    Abcam user community

    Verified customer

    Submitted Nov 05 2014

    Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
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