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Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-N myc interactor/NMI antibody [EPR11065(2)] (ab183724)

Submit a review Submit a question References (7)
Western blot - Anti-N myc interactor/NMI antibody [EPR11065(2)] (ab183724)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-N myc interactor/NMI antibody [EPR11065(2)] (ab183724)
  • Western blot - Anti-N myc interactor/NMI antibody [EPR11065(2)] (ab183724)
  • Western blot - Anti-N myc interactor/NMI antibody [EPR11065(2)] (ab183724)
  • Immunocytochemistry/ Immunofluorescence - Anti-N myc interactor/NMI antibody [EPR11065(2)] (ab183724)
  • Immunoprecipitation - Anti-N myc interactor/NMI antibody [EPR11065(2)] (ab183724)
  • Anti-N myc interactor/NMI antibody [EPR11065(2)] (ab183724)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR11065(2)] to N myc interactor/NMI
  • Suitable for: WB, IHC-P, ICC/IF, IP
  • Knockout validated
  • Reacts with: Human

Conjugates logo Related conjugates and formulations

Carrier Free

Overview

  • Product name

    Anti-N myc interactor/NMI antibody [EPR11065(2)]
    See all N myc interactor/NMI primary antibodies

  • Description

    Rabbit monoclonal [EPR11065(2)] to N myc interactor/NMI

  • Host species

    Rabbit

  • Tested applications

    Suitable for: WB, IHC-P, ICC/IF, IPmore details

  • Species reactivity

    Reacts with: Human

  • Immunogen

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

  • Positive control

    • WB: HeLa, A549, K562 and HepG2 cell lysates. IHC-P: Human tonsil tissue. ICC: HeLa 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

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab183724 in the following tested applications.

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

Application Abreviews Notes
WB
1/10000 - 1/50000. Detects a band of approximately 38 kDa (predicted molecular weight: 35 kDa).
IHC-P
1/50. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
ICC/IF
1/100.
IP
1/40 - 1/60.
Notes
WB
1/10000 - 1/50000. Detects a band of approximately 38 kDa (predicted molecular weight: 35 kDa).
IHC-P
1/50. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
ICC/IF
1/100.
IP
1/40 - 1/60.

Target

  • Relevance

    NMYC interactor (NMI) encodes a protein that interacts with NMYC and CMYC (two members of the oncogene Myc family), and other transcription factors containing a Zip, HLH, or HLH Zip motif. The NMI protein also interacts with all STATs except STAT2 and augments STAT mediated transcription in response to cytokines IL2 and IFN gamma. The NMI mRNA has high expression in myeloid leukemia cell lines.

  • Cellular localization

    Cytoplasmic

  • Database links

    • Alternative names

      • N myc (and STAT) interactor antibody
      • N myc and STAT interactor antibody
      • N myc interactor antibody
      • NMI antibody
      • NMYC and STAT interactor antibody
      • NMYC interactor antibody
      see all

    Images

    • Western blot - Anti-N myc interactor/NMI antibody [EPR11065(2)] (ab183724)
      Western blot - Anti-N myc interactor/NMI antibody [EPR11065(2)] (ab183724)
      All lanes : Anti-N myc interactor/NMI antibody [EPR11065(2)] (ab183724) at 1/1000 dilution

      Lane 1 : Wild-type A549 (Human lung carcinoma cell line) whole cell lysate
      Lane 2 : NMI knockout A549 (Human lung carcinoma cell line) whole cell lysate
      Lane 3 : HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate
      Lane 4 : K562 (Human chronic myelogenous leukemia lymphoblast cell line ) whole cell lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) at 1/10000 dilution

      Predicted band size: 35 kDa
      Observed band size: 39 kDa why is the actual band size different from the predicted?



      Lanes 1-4: Merged signal (red and green). Green - ab183724 observed at 39 kDa. Red - loading control ab8245 observed at 36 kDa.

       ab183724 Anti-N myc interactor/NMI antibody [EPR11065(2)]  was shown to specifically react with N myc interactor/NMI in wild-type A549 cells. Loss of signal was observed when knockout cell line ab267013 (knockout cell lysate ab258077) was used. Wild-type and N myc interactor/NMI knockout samples were subjected to SDS-PAGE. ab183724 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 dilution and 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.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-N myc interactor/NMI antibody [EPR11065(2)] (ab183724)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-N myc interactor/NMI antibody [EPR11065(2)] (ab183724)

      Immunohistochemical analysis of paraffin-embedded Human tonsil tissue labeling N myc interactor/NMI with ab183724 at 1/50 dilution. The slide is counterstained with Hematoxylin.

      Perform heat mediated antigen retrieval with EDTA buffer pH 9 before commencing with IHC staining protocol.

    • Western blot - Anti-N myc interactor/NMI antibody [EPR11065(2)] (ab183724)
      Western blot - Anti-N myc interactor/NMI antibody [EPR11065(2)] (ab183724)

      Lane 1: Wild-type HAP1 cell lysate (20 µg)
      Lane 2:       N myc interactor/NMI knockout HAP1 cell lysate (20 µg)
      Lane 3: HeLa cell lysate (20 µg)
      Lane 4: K562 cell lysate (20 µg)
      Lanes 1 - 4:       Merged signal (red and green). Green - ab183724 observed at 39 kDa. Red - loading control, ab18058, observed at 124 kDa.
      ab183724 was shown to specifically react with N myc interactor/NMI when N myc interactor/NMI knockout samples were used. Wild-type and N myc interactor/NMI knockout samples were subjected to SDS-PAGE. ab183724 and ab18058 (loading control to Vinculin) were diluted at 1/2000 and 1/10 000 respectively and incubated overnight at 4°C. 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 1/10000 dilution for 1 hour at room temperature before imaging.

    • Western blot - Anti-N myc interactor/NMI antibody [EPR11065(2)] (ab183724)
      Western blot - Anti-N myc interactor/NMI antibody [EPR11065(2)] (ab183724)
      All lanes : Anti-N myc interactor/NMI antibody [EPR11065(2)] (ab183724) at 1/20000 dilution

      Lane 1 : K562 cell lysate
      Lane 2 : HeLa cell lysate
      Lane 3 : HepG2 cell lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

      Predicted band size: 35 kDa

    • Immunocytochemistry/ Immunofluorescence - Anti-N myc interactor/NMI antibody [EPR11065(2)] (ab183724)
      Immunocytochemistry/ Immunofluorescence - Anti-N myc interactor/NMI antibody [EPR11065(2)] (ab183724)

      Immunofluorescence analysis of acetone-fixed HeLa cells labeling N myc interactor/NMI with ab183724 at 1/100 dilution. Goat anti-rabbit IgG (Alexa Fluor® 488) at 1/200 dilution was used as the secondary antibody (green). The slide on the right is stained with Dapi (blue).

    • Immunoprecipitation - Anti-N myc interactor/NMI antibody [EPR11065(2)] (ab183724)
      Immunoprecipitation - Anti-N myc interactor/NMI antibody [EPR11065(2)] (ab183724)

      Western blot analysis of K562 cell lysate precipitated with ab183724 at 1/50 dilution. A Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated secondary antibody at 1/1000 dilution was then used. The blocking buffer and dilution buffer was 5% NFDM/TBST.

    • Anti-N myc interactor/NMI antibody [EPR11065(2)] (ab183724)
      Anti-N myc interactor/NMI antibody [EPR11065(2)] (ab183724)

    References (7)

    Publishing research using ab183724? Please let us know so that we can cite the reference in this datasheet.

    ab183724 has been referenced in 7 publications.

    • Wu S  et al. Downregulation of N-myc Interactor Promotes Cervical Cancer Cells Growth by Activating Stat3 Signaling. Cell Biochem Biophys 79:103-111 (2021). PubMed: 33106998
    • Kerr CH  et al. Dynamic rewiring of the human interactome by interferon signaling. Genome Biol 21:140 (2020). PubMed: 32539747
    • Xiong L  et al. N-myc and STAT interactor correlates with severity and prognosis in acute-on-chronic liver failure of hepatitis B virus. J Gastroenterol Hepatol 34:1800-1808 (2019). PubMed: 30771232
    • Yu F  et al. Multi-marker analysis of genomic annotation on gastric cancer GWAS data from Chinese populations. Gastric Cancer 22:60-68 (2019). PubMed: 29859005
    • Sikorski K  et al. A high-throughput pipeline for validation of antibodies. Nat Methods 15:909-912 (2018). PubMed: 30377371
    • Xia J  et al. Genome-Wide Bimolecular Fluorescence Complementation-Based Proteomic Analysis ofToxoplasma gondiiROP18's Human Interactome Shows Its Key Role in Regulation of Cell Immunity and Apoptosis. Front Immunol 9:61 (2018). PubMed: 29459857
    • Xiahou Z  et al. NMI and IFP35 serve as proinflammatory DAMPs during cellular infection and injury. Nat Commun 8:950 (2017). PubMed: 29038465

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