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RabMAb

Anti-Cleaved PARP1抗体[E51] (ab32064)

概述

  • 产品名称
    Anti-Cleaved PARP1抗体[E51]
    参阅全部 Cleaved PARP1 一抗
  • 描述
    兔单克隆抗体[E51] to Cleaved PARP1
  • 特异性
    This antibody is specific for the p25 cleaved form of human PARP1.
  • 经测试应用
    适用于: WB, IHC-Pmore details
    不适用于: ICC/IF
  • 种属反应性
    与反应: Mouse, Rat, Human
    预测可用于: Chinese hamster
  • 免疫原

    Synthetic peptide within Human Cleaved PARP1 aa 150-250. The exact sequence is proprietary.

  • 阳性对照
    • Jurkat cells.
  • 常规说明

    A trial size is available to purchase for this antibody.

    Produced using Abcam's RabMAb® technology. RabMAb® technology is covered by the following U.S. Patents, No. 5, 675, 063 and/or 7, 429, 487.

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

    This product is a recombinant rabbit monoclonal antibody.

性能

应用

Our Abpromise guarantee covers the use of ab32064 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

应用 Ab评论 说明
WB 1/1000 - 1/10000. Predicted molecular weight: 25 kDa.
IHC-P 1/100.
  • 应用说明
    Is unsuitable for ICC/IF.
  • 靶标

    • 功能
      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.
    • 序列相似性
      Contains 1 BRCT domain.
      Contains 1 PARP alpha-helical domain.
      Contains 1 PARP catalytic domain.
      Contains 2 PARP-type zinc fingers.
    • 翻译后修饰
      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.
    • 细胞定位
      Nucleus. Nucleus, nucleolus. Localizes at sites of DNA damage.
    • Information by UniProt
    • 数据库链接
    • 别名
      • 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

    Anti-Cleaved PARP1 antibody [E51] 图像



    • Predicted band size : 25 kDa

      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 - ab32064 observed at 30 kDa. Red - loading control, ab8245, observed at 37 kDa.

      ab32064 was shown to specifically react with PARP1 when PARP1 knockout samples were used. Wild-type and PARP1 knockout samples were subjected to SDS-PAGE. Ab32064 and ab8245 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 1/10000 dilutions. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) ab216773 and 680CW Goat anti Mouse secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.

    • All lanes : Anti-Cleaved PARP1 antibody [E51] (ab32064) at 1/1000 dilution

      Lane 1 : HeLa treated with 1uM Staurosporine for 3 hours whole cell lysates
      Lane 2 : Untreated HeLa whole cell lysates
      Lane 3 : NIH/3T3 treated with 1uM Staurosporine for 3 hours whole cell lysates
      Lane 4 : Untreated NIH/3T3 whole cell lysates

      Lysates/proteins at 20 µg per lane.

      Secondary
      Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

      Predicted band size : 25 kDa
      Observed band size : 25 kDa

      Blocking/Dilution buffer 5% NFDM/TBST

      Exposure time :
      Lane 1,2: 1 second
      Lane 3,4: 8 seconds

    • Immunohistochemical staining of paraffin embedded rat colon with purified ab32064 at a working dilution of 1/100. The secondary antibody used is a HRP polymer for rabbit IgG. The sample is counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.

    • All lanes : Anti-Cleaved PARP1 antibody [E51] (ab32064) at 1/1000 dilution

      Lane 1 : RAW264.7 cell lysate
      Lane 2 : NIH/3T3 cell lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      HRP goat anti-rabbit (H+L) at 1/1000 dilution

      Predicted band size : 25 kDa
      Observed band size : 25 kDa

      Blocking buffer: 5% NFDM/TBST

      Dilution buffer: 5% NFDM/TBST

    • All lanes : Anti-Cleaved PARP1 antibody [E51] (ab32064) at 1/10000 dilution

      Lane 1 : Untreated Jurkat cell lysate
      Lane 2 : Jurkat cell lysate treated with camptothecin

      Lysates/proteins at 10 µg per lane.

      Secondary
      HRP goat anti-rabbit (H+L) at 1/1000 dilution

      Predicted band size : 25 kDa
      Observed band size : 25 kDa

      Blocking buffer: 5% NFDM/TBST

      Dilution buffer: 5% NFDM/TBST

    • Immunohistochemical staining of paraffin embedded human ovarian carcinoma with purified ab32064 at a working dilution of 1 in 100. The secondary antibody used is a HRP polymer for rabbit IgG. The sample is counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.


    • Developed using the ECL technique

      Performed under reducing conditions.

      Predicted band size : 25 kDa
      Observed band size : 25 kDa


      Exposure time : 10 seconds

      Jurkat cells were incubated at 37°C for 24 hours with vehicle control (0 μM) and different concentrations of 15-Acetoxyscirpenol (ab142381). Increased expression of cleaved PARP1 (ab32064) in Jurkat cells correlates with an increase in 15-Acetoxyscirpenol concentration, as described in literature.

      Whole cell lysates were prepared with RIPA buffer (containing protease inhibitors and sodium orthovanadate), 20μg of each were loaded on the gel and the WB was run under reducing conditions. After transfer the membrane was blocked for an hour using 5% BSA before being incubated with ab32064 at 1/10000 dilution and ab8227 at 1 μg/ml overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP (ab97051) at 1/10000 and visualised using ECL development solution.

    • Anti-Cleaved PARP1 antibody [E51] (ab32064) at 1/50000 dilution + MCF7 cell lysate at 100 µg

      Secondary
      HRP-conjugated Goat anti-rabbit IgG polyclonal at 1/50000 dilution
      Developed using the ECL technique

      Performed under reducing conditions.

      Predicted band size : 25 kDa
      Observed band size : 25 kDa


      Exposure time : 15 minutes

      This image is courtesy of an anonymous Abreview

      See Abreview

    • All lanes : Anti-Cleaved PARP1 antibody [E51] (ab32064) at 1/1000000 dilution

      Lane 1 : Jurkat cell lysate. Untreated.
      Lane 2 : Jurkat cell lysate. Treated with Camptothecin.


      Predicted band size : 25 kDa
      Observed band size : 25 kDa
    • All lanes : Anti-Cleaved PARP1 antibody [E51] (ab32064) at 1/1000 dilution

      Lane 1 : PC-12 treated with 1uM Staurosporine for 3 hours whole cell lysates
      Lane 2 : Untreated PC-12 whole cell lysates

      Lysates/proteins at 20 µg per lane.

      Secondary
      Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

      Predicted band size : 25 kDa
      Observed band size : 25 kDa


      Exposure time : 30 seconds

      Blockinng/Diluting buffer 5% NFDM/TBST

    • Immunohistochemical staining of paraffin embedded human breast carcinoma with unpurified ab32064 at a 1/100 dilution.

    Anti-Cleaved PARP1 antibody [E51] (ab32064)参考文献

    This product has been referenced in:
    • Szobi A  et al. Analysis of necroptotic proteins in failing human hearts. J Transl Med 15:86 (2017). WB ; Human . Read more (PubMed: 28454582) »
    • Özata DM  et al. Loss of miR-514a-3p regulation of PEG3 activates the NF-kappa B pathway in human testicular germ cell tumors. Cell Death Dis 8:e2759 (2017). WB ; Human . Read more (PubMed: 28471449) »

    See all 29 Publications for this product

    Product Wall

    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application
    Western blot
    Sample
    Mouse Tissue lysate - whole (Brain)
    Gel Running Conditions
    Reduced Denaturing (4-20%)
    Loading amount
    40 µg
    Specification
    Brain
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
    Username

    Dr. SAIFUDEEN ISMAEL

    Verified customer

    提交于 Jun 06 2017

    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (U2OS cells)
    Gel Running Conditions
    Reduced Denaturing (4-12% Gradient Gel)
    Loading amount
    20 µg
    Specification
    U2OS cells
    Blocking step
    Licor Blocking Buffer as blocking agent for 30 minute(s) · Concentration: 100% · Temperature: 25°C
    Username

    Abcam user community

    Verified customer

    提交于 Mar 02 2017

    Application
    Western blot
    Sample
    Zebrafish Tissue lysate - whole (whole embryo)
    Gel Running Conditions
    Reduced Denaturing (12%)
    Loading amount
    10 µg
    Treatment
    25uM tamoxifen
    Specification
    whole embryo
    Blocking step
    BSA as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
    Username

    Abcam user community

    Verified customer

    提交于 Jul 28 2015

    Application
    Immunocytochemistry
    Sample
    Human Cultured Cells (Ovarian cancer cell line)
    Permeabilization
    Yes - 0.25% tritonx X-100
    Specification
    Ovarian cancer cell line
    Blocking step
    Background buster as blocking agent for 30 minute(s) · Concentration: 100% · Temperature: 25°C
    Fixative
    Formaldehyde
    Username

    Abcam user community

    Verified customer

    提交于 Aug 11 2014

    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Mouse Tissue sections (Mice pancreas)
    Antigen retrieval step
    Other
    Permeabilization
    No
    Specification
    Mice pancreas
    Blocking step
    No blocking step used for 30 minute(s) · Concentration: 100% · Temperature: 25°C
    Fixative
    Paraformaldehyde
    Username

    Abcam user community

    Verified customer

    提交于 Aug 11 2014

    The antigen retrieval was heat-mediated with citrate buffer, pH 6.0. The laboratory uses a pressure cooker but a microwave, steamer, or waterbath should work too.

    Application
    Flow Cytometry
    Sample
    Human Cell (MCF7)
    Permeabilization
    Yes - E BIOSCIENCES PERMEABILISATION BUFFER
    Gating Strategy
    CD44 POSITIVE
    Specification
    MCF7
    Preparation
    Cell harvesting/tissue preparation method: single cell suspension
    Sample buffer: 0.1% PBS BSA
    Fixation
    Formaldehyde
    Username

    Abcam user community

    Verified customer

    提交于 Dec 06 2013


    The concentration for the aforementioned lot is 0.3770 mg/ml

    Thank you for contacting Abcam regarding ab32064.


    I have confirmed with the laboratory that we currently do not have any data demonstrating the use of this antibody with a fluorescently conjugated secondary antibody. However, we expect ...

    Read More
    Abcam has not validated the combination of species/application used in this Abreview.
    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Rat Cell (cell line IEC-6)
    Permeabilization
    Yes - 0.1% Triton X-100
    Specification
    cell line IEC-6
    Blocking step
    BSA as blocking agent for 30 minute(s) · Concentration: 1% · Temperature: Rm°C
    Fixative
    Formaldehyde
    Username

    Abcam user community

    Verified customer

    提交于 Jan 13 2012

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