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使用敲除细胞株进行验证重组RabMAb

重组Anti-CDKN2A/p14ARF抗体[EPR17878] - BSA and Azide free (ab224273)

Submit a review Submit a question References (1)
Immunocytochemistry/ Immunofluorescence - Anti-CDKN2A/p14ARF antibody [EPR17878] - BSA and Azide free (ab224273)
  • Flow Cytometry (Intracellular) - Anti-CDKN2A/p14ARF antibody [EPR17878] - BSA and Azide free (ab224273)
  • Immunoprecipitation - Anti-CDKN2A/p14ARF antibody [EPR17878] - BSA and Azide free (ab224273)
  • Western blot - Anti-CDKN2A/p14ARF antibody [EPR17878] - BSA and Azide free (ab224273)
  • Immunocytochemistry/ Immunofluorescence - Anti-CDKN2A/p14ARF antibody [EPR17878] - BSA and Azide free (ab224273)
  • Anti-CDKN2A/p14ARF antibody [EPR17878] - BSA and Azide free (ab224273)

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 - BSA and Azide free
  • Suitable for: IP, ICC/IF, WB, Flow Cyt (Intra)
  • Knockout validated
  • Reacts with: Human

Conjugates logo Related conjugates and formulations

Alexa Fluor® 488 Alexa Fluor® 555 Alexa Fluor® 594 Alexa Fluor® 647 APC PE Unconjugated

概述

  • 产品名称

    Anti-CDKN2A/p14ARF抗体[EPR17878] - BSA and Azide free
    参阅全部 CDKN2A/p14ARF 一抗

  • 描述

    兔单克隆抗体[EPR17878] to CDKN2A/p14ARF - BSA and Azide free

  • 宿主

    Rabbit

  • 经测试应用

    适用于: IP, ICC/IF, WB, Flow Cyt (Intra)more details

  • 种属反应性

    与反应: Human

  • 免疫原

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

  • 阳性对照

    • 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.

  • 常规说明

    ab224273 is the carrier-free version of ab185620.

    Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.

    This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.

    Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

    This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.

    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.

性能

应用

The Abpromise guarantee

Abpromise™承诺保证使用ab224273于以下的经测试应用

“应用说明”部分 下显示的仅为推荐的起始稀释度；实际最佳的稀释度/浓度应由使用者检定。

应用 Ab评论 说明
IP
Use at an assay dependent concentration.
ICC/IF
Use at an assay dependent concentration.
WB
Use at an assay dependent concentration. Detects a band of approximately 14 kDa (predicted molecular weight: 14 kDa).
Flow Cyt (Intra)
Use at an assay dependent concentration.

ab199376 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
说明
IP
Use at an assay dependent concentration.
ICC/IF
Use at an assay dependent concentration.
WB
Use at an assay dependent concentration. Detects a band of approximately 14 kDa (predicted molecular weight: 14 kDa).
Flow Cyt (Intra)
Use at an assay dependent concentration.

ab199376 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

靶标

  • 相关性

    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.

  • 细胞定位

    Cytoplasmic and Nuclear

  • 数据库链接

    • 别名

      • 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

    图片

    • Immunocytochemistry/ Immunofluorescence - Anti-CDKN2A/p14ARF antibody [EPR17878] - BSA and Azide free (ab224273)
      Immunocytochemistry/ Immunofluorescence - Anti-CDKN2A/p14ARF antibody [EPR17878] - BSA and Azide free (ab224273)

      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.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab185620).

    • Flow Cytometry (Intracellular) - Anti-CDKN2A/p14ARF antibody [EPR17878] - BSA and Azide free (ab224273)
      Flow Cytometry (Intracellular) - Anti-CDKN2A/p14ARF antibody [EPR17878] - BSA and Azide free (ab224273)

       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. 

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab185620). 

       

       

    • Immunoprecipitation - Anti-CDKN2A/p14ARF antibody [EPR17878] - BSA and Azide free (ab224273)
      Immunoprecipitation - Anti-CDKN2A/p14ARF antibody [EPR17878] - BSA and Azide free (ab224273)

      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.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab185620).

    • Western blot - Anti-CDKN2A/p14ARF antibody [EPR17878] - BSA and Azide free (ab224273)
      Western blot - Anti-CDKN2A/p14ARF antibody [EPR17878] - BSA and Azide free (ab224273)
      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.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab185620).

    • Immunocytochemistry/ Immunofluorescence - Anti-CDKN2A/p14ARF antibody [EPR17878] - BSA and Azide free (ab224273)
      Immunocytochemistry/ Immunofluorescence - Anti-CDKN2A/p14ARF antibody [EPR17878] - BSA and Azide free (ab224273)

      This ICC data was generated using the same anti-CDKN2A/p14ARF antibody clone [EPR17878] in a different buffer formulation (cat# 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.

    • Anti-CDKN2A/p14ARF antibody [EPR17878] - BSA and Azide free (ab224273)
      Anti-CDKN2A/p14ARF antibody [EPR17878] - BSA and Azide free (ab224273)

    文献 (1)

    发表研究结果有使用 ab224273？请让我们知道，以便我们可以引用本数据表中的参考文章。

    ab224273 被引用在 1 文献中.

    • Coleman KE  et al. SENP8 limits aberrant neddylation of NEDD8 pathway components to promote cullin-RING ubiquitin ligase function. Elife 6: (2017). PubMed: 28475037

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