Astana Biomed Group, an authorized Abcam distributor in Central Asia

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: WB, Flow Cyt, IP
  • Knockout validated
  • Reacts with: Human

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 & Species

    Application Species
    Flow Cyt
    Human
    IP
    Human
    WB
    Human
    See all applications and species data

  • 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

  • General notes

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

     

    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.

Properties

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

    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).

  • 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

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

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
For licensing inquiries, please contact partnerships@abcam.com

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