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遺伝子ノックアウト細胞株 検証済リコンビナントRabMAb

Anti-CDKN2A/p14ARF 抗体 [EPR17878] (ab185620)

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Western blot - Anti-CDKN2A/p14ARF antibody [EPR17878] (ab185620)
  • Western blot - Anti-CDKN2A/p14ARF antibody [EPR17878] (ab185620)
  • Western blot - Anti-CDKN2A/p14ARF antibody [EPR17878] (ab185620)
  • Immunocytochemistry/ Immunofluorescence - Anti-CDKN2A/p14ARF antibody [EPR17878] (ab185620)
  • Flow Cytometry (Intracellular) - Anti-CDKN2A/p14ARF antibody [EPR17878] (ab185620)
  • Immunoprecipitation - Anti-CDKN2A/p14ARF antibody [EPR17878] (ab185620)
  • Anti-CDKN2A/p14ARF antibody [EPR17878] (ab185620)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR17878] to CDKN2A/p14ARF
  • Suitable for: Flow Cyt (Intra), ICC/IF, IP, WB
  • Knockout validated
  • Reacts with: Human

Conjugates logo Related conjugates and formulations

Alexa Fluor® 488 Alexa Fluor® 555 Alexa Fluor® 594 Alexa Fluor® 647 APC Carrier Free PE

製品の概要

  • 製品名

    Anti-CDKN2A/p14ARF antibody [EPR17878]
    CDKN2A/p14ARF 一次抗体 製品一覧

  • 製品の詳細

    Rabbit monoclonal [EPR17878] to CDKN2A/p14ARF

  • 由来種

    Rabbit

  • アプリケーション

    適用あり: Flow Cyt (Intra), ICC/IF, IP, WBmore details

  • 種交差性

    交差種: Human

  • 免疫原

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

  • ポジティブ･コントロール

    • WB: HeLa whole cell lysate transfected with CDKN2A/p14ARF with His-tag; PC-3 and HEK-293 whole cell lysates. ICC/IF: HeLa cells transfected with CDKN2A/p14ARF. Flow Cyt (intra): HeLa cells transfected with CDKN2A/p14ARF-GFP. IP: CDKN2A/p14ARF transfected HeLa whole cell lysate.

  • 特記事項

    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.

製品の特性

アプリケーション

The Abpromise guarantee

Abpromise保証は、 次のテスト済みアプリケーションにおけるab185620の使用に適用されます

アプリケーションノートには、推奨の開始希釈率がありますが、適切な希釈率につきましてはご検討ください。

アプリケーション Abreviews 特記事項
Flow Cyt (Intra)
1/120.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
ICC/IF
1/250.
IP
1/80.
WB
1/1000. Detects a band of approximately 14 kDa (predicted molecular weight: 14 kDa).
特記事項
Flow Cyt (Intra)
1/120.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
ICC/IF
1/250.
IP
1/80.
WB
1/1000. Detects a band of approximately 14 kDa (predicted molecular weight: 14 kDa).

ターゲット情報

  • 関連性

    The gene for CDK2NA generates several transcripts/proteins which differ from each other in their first exons. Three of these transcripts are generated by alternative splicing (isoform 1 a.k.a p16INK4A, isoform 2 and isoform 3 a.k.a p12), two of which are known to function as inhibitors of CDK4 kinase. One other transcript that is generated from this gene contains an alternate reading frame (ARF), with the first exon located 20kb upstream of the remainder of the gene(isoform 4 a.k.a. p14ARF, p19ARF, ARF). In spite of the structural and some functional differences, all the proteins encoded by the CDKN2A gene are involved in cell cycle G1 control.

  • 細胞内局在

    Cytoplasmic and Nuclear

  • 参照データベース

    • 別名

      • ARF antibody
      • CDK4 inhibitor p16 INK4 antibody
      • CDK4I antibody
      • CDKN2 antibody
      • CDKN2A antibody
      • Cell cycle negative regulator beta antibody
      • CMM2 antibody
      • Cyclin dependent kinase 4 inhibitor A antibody
      • Cyclin dependent kinase inhibitor 2A antibody
      • INK4 antibody
      • INK4a antibody
      • Melanoma p16 inhibits CDK4 antibody
      • MLM antibody
      • MTS 1 antibody
      • MTS1 antibody
      • Multiple tumor suppressor 1 antibody
      • p14 antibody
      • p16 antibody
      • p16 INK4a antibody
      • p16INK4a antibody
      • p19 antibody
      • P19ARF antibody
      • TP16 antibody
      see all

    画像

    • Western blot - Anti-CDKN2A/p14ARF antibody [EPR17878] (ab185620)
      Western blot - Anti-CDKN2A/p14ARF antibody [EPR17878] (ab185620)
      All lanes : Anti-CDKN2A/p14ARF antibody [EPR17878] (ab185620) at 1/1000 dilution

      Lane 1 : Wild-type HeLa whole cell lysate
      Lane 2 : CDKN2A knockout HeLa whole cell lysate

      Lysates/proteins at 20 µg per lane.

      Predicted band size: 14 kDa



      Lanes 1 - 2: Merged signal (red and green). Green - ab185620 observed at 14 kDa. Red - loading control, ab9484, observed at 37 kDa.

      ab185620 was shown to specifically react with CDKN2A/p14ARF in wild-type HeLa cells as signal was lost in CDKN2A knockout cells. Wild-type and CDKN2A knockout samples were subjected to SDS-PAGE. Ab185620 and ab9484 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/20000 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 1/20000 dilution for 1 hour at room temperature before imaging.

    • Western blot - Anti-CDKN2A/p14ARF antibody [EPR17878] (ab185620)
      Western blot - Anti-CDKN2A/p14ARF antibody [EPR17878] (ab185620)
      All lanes : Anti-CDKN2A/p14ARF antibody [EPR17878] (ab185620) at 1/5000 dilution

      Lane 1 : HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate transfected with empty vector (vector control)
      Lane 2 : HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate transfected with CDKN2A/p14ARF with His-tag

      Lysates/proteins at 10 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

      Predicted band size: 14 kDa


      Exposure time: 1 second


      Blocking/Dilution buffer: 5% NFDM/TBST.

    • Western blot - Anti-CDKN2A/p14ARF antibody [EPR17878] (ab185620)
      Western blot - Anti-CDKN2A/p14ARF antibody [EPR17878] (ab185620)
      All lanes : Anti-CDKN2A/p14ARF antibody [EPR17878] (ab185620) at 1/1000 dilution

      Lane 1 : PC-3 (Human prostate adenocarcinoma cell line) whole cell lysate
      Lane 2 : HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

      Predicted band size: 14 kDa
      Observed band size: 14 kDa


      Exposure time: 3 minutes


      Blocking/Dilution buffer: 5% NFDM/TBST.

      The expression level of p14 ARF is relatively low in most cell types and will be induced by oncogene expression. (PMID: 9694807; PMID: 9694806).

    • Immunocytochemistry/ Immunofluorescence - Anti-CDKN2A/p14ARF antibody [EPR17878] (ab185620)
      Immunocytochemistry/ Immunofluorescence - Anti-CDKN2A/p14ARF antibody [EPR17878] (ab185620)

      Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (human cervix adenocarcinoma) cells transfected with CDKN2A/p14ARF or empty vector, labeling CDKN2A/p14ARF with ab185620 at 1/250 dilution, followed by Goat Anti-Rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution 

      The nuclear counterstain is DAPI (blue).

      Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) at 1/1000 dilution and Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed (ab150120) at 1/1000 dilution (red).

      The negative controls are as follows:-

      -ve control 1: ab185620 at 1/250 dilution followed by ab150120 at 1/1000 dilution.

      -ve control 2: ab7291 at 1/1000 dilution followed by ab150077  at 1/1000 dilution.

    • Flow Cytometry (Intracellular) - Anti-CDKN2A/p14ARF antibody [EPR17878] (ab185620)
      Flow Cytometry (Intracellular) - Anti-CDKN2A/p14ARF antibody [EPR17878] (ab185620)

      Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed HeLa (Human epithelial cell line from cervix adenocarcinoma) cells transfected with CDKN2A/p14ARF-GFP labeling CDKN2A/p14ARF with ab185620 at 1/120 dilution (red) compared with a Rabbit IgG,monoclonal [EPR25A] - Isotype Control (ab172730) (black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody; blue). Goat anti Rabbit IgG (PE) at 1/150 dilution was used as the secondary antibody. 

       

       

    • Immunoprecipitation - Anti-CDKN2A/p14ARF antibody [EPR17878] (ab185620)
      Immunoprecipitation - Anti-CDKN2A/p14ARF antibody [EPR17878] (ab185620)

      CDKN2A/p14ARF was immunoprecipitated from 1mg of CDKN2A/p14ARF transfected HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell lysate with ab185620 at 1/80 dilution.

      Western blot was performed from the immunoprecipitate using ab185620 at 1/1000 dilution.

      VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.

      Lane 1: CDKN2A/p14ARF transfected HeLa whole cell lysate, 10µg (Input).

      Lane 2: ab185620 IP in CDKN2A/p14ARF transfected HeLa whole cell lysate.

      Lane 3: Rabbit IgG,monoclonal[EPR25A] - Isotype Control (ab172730) instead of ab185620 in CDKN2A/p14ARF transfected HeLa whole cell lysate.

      Blocking and dilution buffer and concentration: 5% NFDM/TBST.

      Exposure time: 1 second.

    • Anti-CDKN2A/p14ARF antibody [EPR17878] (ab185620)
      Anti-CDKN2A/p14ARF antibody [EPR17878] (ab185620)

    参考文献 (5)

    ab185620 を使用した論文を発表された方は、こちらまでお知らせください。データシートに掲載させていただきます。

    ab185620 は 5 報の論文で使用されています。

    • Liang F  et al. Long noncoding RNA LINC01088 inhibits esophageal squamous cell carcinoma progression by targeting the NPM1-HDM2-p53 axis. Acta Biochim Biophys Sin (Shanghai) 55:367-381 (2023). PubMed: 36942988
    • Shi WK  et al. The Cell Cycle-Associated Protein CDKN2A May Promotes Colorectal Cancer Cell Metastasis by Inducing Epithelial-Mesenchymal Transition. Front Oncol 12:834235 (2022). PubMed: 35311137
    • Miao J  et al. Klotho retards renal fibrosis through targeting mitochondrial dysfunction and cellular senescence in renal tubular cells. Physiol Rep 9:e14696 (2021). PubMed: 33463897
    • Horvat A  et al. Helicobacter pylori pathogen regulates p14ARF tumor suppressor and autophagy in gastric epithelial cells. Oncogene 37:5054-5065 (2018). PubMed: 29849123
    • Coleman KE  et al. SENP8 limits aberrant neddylation of NEDD8 pathway components to promote cullin-RING ubiquitin ligase function. Elife 6:N/A (2017). PubMed: 28475037

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