製品の概要

  • 製品名Anti-Cleaved PARP antibody [E51]
    Cleaved PARP 一次抗体 製品一覧
  • 製品の詳細
    Rabbit monoclonal [E51] to Cleaved PARP
  • 特異性This antibody is specific for the p25 cleaved form of human PARP.
  • アプリケーション適用あり: WB, IHC-Pmore details
    適用なし: ICC/IF
  • 種交差性
    交差種: Mouse, Rat, Human
    交差が予測される動物種: Chinese Hamster
  • 免疫原

    Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) corresponding to Human Cleaved PARP aa 150-250.

  • ポジティブ・コントロール
    • Jurkat cells.
  • 特記事項

    This product is a recombinant rabbit monoclonal antibody.

    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.

    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.

    A trial size is available to purchase for this antibody.

     

    Alternative versions available:

    Anti-Cleaved PARP antibody (HRP) [E51] (ab194217)

製品の特性

アプリケーション

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.

アプリケーション Abreviews 特記事項
WB 1/1000 - 1/10000. Predicted molecular weight: 25 kDa.
IHC-P 1/100.
  • 追加情報Is unsuitable for ICC/IF.
  • ターゲット情報

    • 機能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.
    • 配列類似性Contains 1 BRCT domain.
      Contains 1 PARP alpha-helical domain.
      Contains 1 PARP catalytic domain.
      Contains 2 PARP-type zinc fingers.
    • 翻訳後修飾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.
    • 細胞内局在Nucleus. Nucleus, nucleolus. Localizes at sites of DNA damage.
    • Information by UniProt
    • 参照データベース
    • 別名
      • 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 PARP antibody [E51] 画像



    • 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 PARP 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 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.

    • All lanes : Anti-Cleaved PARP 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 PARP 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 PARP (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 PARP 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 PARP 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 PARP 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 PARP antibody [E51] (ab32064) 使用論文

    This product has been referenced in:
    • Qin Q  et al. miR-134 inhibits non-small cell lung cancer growth by targeting the epidermal growth factor receptor. J Cell Mol Med N/A:N/A (2016). IHC . Read more (PubMed: 27241841) »
    • Sanchez-Niño MD  et al. Albumin-induced apoptosis of tubular cells is modulated by BASP1. Cell Death Dis 6:e1644 (2015). WB ; Human . Read more (PubMed: 25675304) »

    See all 21 Publications for this product

    Product Wall

    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
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    投稿 Jul 28 2015

    Application Immunocytochemistry
    Blocking step Background buster as blocking agent for 30 minute(s) · Concentration: 100% · Temperature: 25°C
    Sample Human Cultured Cells (Ovarian cancer cell line)
    Specification Ovarian cancer cell line
    Permeabilization Yes - 0.25% tritonx X-100
    Fixative Formaldehyde
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    投稿 Aug 11 2014

    Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Blocking step No blocking step used for 30 minute(s) · Concentration: 100% · Temperature: 25°C
    Antigen retrieval step Other
    Sample Mouse Tissue sections (Mice pancreas)
    Specification Mice pancreas
    Permeabilization No
    Fixative Paraformaldehyde
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    投稿 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
    Fixation Formaldehyde
    Permeabilization Yes - E BIOSCIENCES PERMEABILISATION BUFFER
    Sample Human Cell (MCF7)
    Specification MCF7
    Gating Strategy CD44 POSITIVE
    Preparation Cell harvesting/tissue preparation method: single cell suspension
    Sample buffer: 0.1% PBS BSA
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    投稿 Dec 06 2013

    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application Western blot
    Loading amount 50 µg
    Gel Running Conditions Reduced Denaturing (10)
    Sample Human Cell lysate - whole cell (MCF7)
    Specification MCF7
    Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 37°C
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    投稿 Sep 09 2013

    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application Immunoprecipitation
    Immuno-precipitation step Staph. aureus
    Sample Human Cell lysate - whole cell (MCF7)
    Specification MCF7
    Total protein in input 100 µg
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    投稿 Sep 06 2013

    Abcam has not validated the combination of species/application used in this Abreview.
    Application Immunoprecipitation
    Sample Human Cell lysate - whole cell (MCF7)
    Total protein in input 50 µg
    Treatment T3 100 micro moles for 48 hrs
    Immuno-precipitation step Staph. aureus
    Specification MCF7
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    投稿 Jun 27 2013

    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application Western blot
    Loading amount 100 µg
    Gel Running Conditions Reduced Denaturing
    Sample Human Cell lysate - whole cell (MCF7)
    Specification MCF7
    Treatment T3 100 micro moles for 48 hrs
    Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 37°C
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    投稿 Jun 27 2013


    The concentration for the aforementioned lot is 0.3770 mg/ml

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