Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-PARP1 antibody [E102] (ab32138)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [E102] to PARP1
  • Suitable for: WB, IHC-P, Flow Cyt
  • Knockout validated
  • Reacts with: Human

Overview

  • Product name

    Anti-PARP1 antibody [E102]
    See all PARP1 primary antibodies

  • Description

    Rabbit monoclonal [E102] to PARP1

  • Host species

    Rabbit

  • Specificity

    ab32138 recognises both pro-form and p25 cleaved form of PARP1.

  • Tested Applications & Species

    Application Species
    Flow Cyt
    Human
    IHC-P
    Human
    WB
    Human
    See all applications and species data

  • Immunogen

    Synthetic peptide within Human PARP1 (N terminal). The exact sequence is proprietary.

  • Positive control

    • WB: Wild type HAP1 whole cell lysate; HeLa whole cell lysate (ab150035); Jurkat whole cell lysate (ab7899); HEK-293T cell lysate. IHC-P: Human brain tissue. ICC/IF: HeLa cells. Flow Cyt: Jurkat 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.

Properties

Images

  • Western blot - Anti-PARP1 antibody [E102] (ab32138)
    Western blot - Anti-PARP1 antibody [E102] (ab32138)

    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 - ab32138 observed at 125 kDa. Red - loading control, ab8245, observed at 37 kDa.

    ab32138 was shown to specifically react with PARP1 when PARP1 knockout samples were used. Wild-type and PARP1 knockout samples were subjected to SDS-PAGE. ab32138 and ab8245 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/10 000 dilution respectively. Blots were developed with 800CW Goat anti Rabbit and 680CW Goat anti Mouse secondary antibodies at 1/10 000 dilution for 1 hour at room temperature before imaging.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PARP1 antibody [E102] (ab32138)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PARP1 antibody [E102] (ab32138)

    Immunohistochemical analysis of PARP1 expression in paraffin embedded human brain tissue section, using 1/25 ab32138.

    Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

  • Flow Cytometry - Anti-PARP1 antibody [E102] (ab32138)
    Flow Cytometry - Anti-PARP1 antibody [E102] (ab32138)

    Overlay histogram showing Jurkat (human T cell leukemia cell line from peripheral blood) cells stained with ab32138 (red line). The cells were fixed with methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab32138, 1/50 dilution) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1:500 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit monoclonal IgG (0.5µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a decreased signal in Jurkat cells fixed with 4% paraformaldehyde (10 min)/permeabilized in 0.1% PBS-Tween used under the same conditions.

  • Western blot - Anti-PARP1 antibody [E102] (ab32138)
    Western blot - Anti-PARP1 antibody [E102] (ab32138) Image uploaded from Edigene testing
    All lanes : Anti-PARP1 antibody [E102] (ab32138) at 1/1000 dilution

    Lane 1 : Wild-type HEK-293T cell lysate
    Lane 2 : PARP1 knockout HEK-293T cell lysate

    Lysates/proteins at 20 µg per lane.

    Performed under reducing conditions.

    Predicted band size: 113 kDa
    Observed band size: 113 kDa



    Lanes 1- 2: Merged signal (red and green). Green - ab32138 observed at 113 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.

     ab32138 was shown to react with PARP1 in wild-type HEK-293T cells in western blot. Loss of signal was observed when knockout cell line ab266598 (knockout cell lysate ab257017) was used. Wild-type HEK-293T and PARP1 knockout HEK-293T cell lysates were subjected to SDS-PAGE. ab32138 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) overnight at 4°C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye®800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye®680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

  • Western blot - Anti-PARP1 antibody [E102] (ab32138)
    Western blot - Anti-PARP1 antibody [E102] (ab32138)
    All lanes : Anti-PARP1 antibody [E102] (ab32138) at 1/1000 dilution

    Lane 1 : Jurkat (human T cell leukemia cell line from peripheral blood) cell lysate
    Lane 2 : Jurkat + Camptothecin cell lysate

    Predicted band size: 113 kDa
    Observed band size: 120,25 kDa
    why is the actual band size different from the predicted?

  • Anti-PARP1 antibody [E102] (ab32138)
    Anti-PARP1 antibody [E102] (ab32138)

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