Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-SMYD3 antibody [EPR11106(2)] - N-terminal (ab183498)

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Western blot - Anti-SMYD3 antibody [EPR11106(2)] - N-terminal (ab183498)
  • Western blot - Anti-SMYD3 antibody [EPR11106(2)] - N-terminal (ab183498)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SMYD3 antibody [EPR11106(2)] - N-terminal (ab183498)
  • Immunocytochemistry/ Immunofluorescence - Anti-SMYD3 antibody [EPR11106(2)] - N-terminal (ab183498)
  • Immunoprecipitation - Anti-SMYD3 antibody [EPR11106(2)] - N-terminal (ab183498)
  • Flow Cytometry (Intracellular) - Anti-SMYD3 antibody [EPR11106(2)] - N-terminal (ab183498)
  • Western blot - Anti-SMYD3 antibody [EPR11106(2)] - N-terminal (ab183498)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SMYD3 antibody [EPR11106(2)] - N-terminal (ab183498)
  • Anti-SMYD3 antibody [EPR11106(2)] - N-terminal (ab183498)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR11106(2)] to SMYD3 - N-terminal
  • Suitable for: Flow Cyt (Intra), IHC-P, WB, ICC/IF, IP
  • Knockout validated
  • Reacts with: Human

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Carrier Free

Overview

  • Product name

    Anti-SMYD3 antibody [EPR11106(2)] - N-terminal
    See all SMYD3 primary antibodies

  • Description

    Rabbit monoclonal [EPR11106(2)] to SMYD3 - N-terminal

  • Host species

    Rabbit

  • Tested applications

    Suitable for: Flow Cyt (Intra), IHC-P, WB, 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, HAP1, 293, MCF-7 and T47-D cell lysates. IHC-P: Human testis and skeletal muscle tissue. ICC: HeLa cells. Flow Cyt (intra): 293 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 ab183498 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
Flow Cyt (Intra)
Use at an assay dependent concentration.
IHC-P
1/50 - 1/100. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
WB
1/1000 - 1/10000. Detects a band of approximately 45 kDa (predicted molecular weight: 49 kDa).
ICC/IF
1/500.
IP
1/40 - 1/60.
Notes
Flow Cyt (Intra)
Use at an assay dependent concentration.
IHC-P
1/50 - 1/100. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
WB
1/1000 - 1/10000. Detects a band of approximately 45 kDa (predicted molecular weight: 49 kDa).
ICC/IF
1/500.
IP
1/40 - 1/60.

Target

  • Function

    Histone methyltransferase. Specifically methylates 'Lys-4' of histone H3, inducing di- and tri-methylation, but not monomethylation. Plays an important role in transcriptional activation as a member of an RNA polymerase complex. Binds DNA containing 5'-CCCTCC-3' or 5'-GAGGGG-3' sequences.

  • Tissue specificity

    Expressed in skeletal muscles and testis. Overexpressed in a majority of colorectal and hepatocellular carcinomas.

  • Sequence similarities

    Belongs to the histone-lysine methyltransferase family.
    Contains 1 MYND-type zinc finger.
    Contains 1 SET domain.

  • Cellular localization

    Cytoplasm. Nucleus. Mainly cytoplasmic when cells are arrested at G0/G1. Accumulates in the nucleus at S phase and G2/M.
  • Information by UniProt

  • Database links

    • Alternative names

      • bA74P14.1 (novel protein) antibody
      • bA74P14.1 antibody
      • FLJ21080 antibody
      • histone lysine N methyltransferase SMYD3 antibody
      • KMT3E antibody
      • MGC104324 antibody
      • SET and MYND domain containing 3 antibody
      • SET and MYND domain containing protein 3 antibody
      • SET and MYND domain-containing protein 3 antibody
      • SMYD 3 antibody
      • Smyd3 antibody
      • SMYD3 protein antibody
      • SMYD3_HUMAN antibody
      • Zinc finger MYND domain containing 1 antibody
      • Zinc finger MYND domain containing protein 1 antibody
      • Zinc finger MYND domain-containing protein 1 antibody
      • Zinc finger protein subfamily 3A MYND domain containing 1 antibody
      • Zinc finger protein, subfamily 3A (MYND domain containing), 1 antibody
      • ZMYND 1 antibody
      • ZMYND1 antibody
      • ZNFN3A1 antibody
      see all

    Images

    • Western blot - Anti-SMYD3 antibody [EPR11106(2)] - N-terminal (ab183498)
      Western blot - Anti-SMYD3 antibody [EPR11106(2)] - N-terminal (ab183498)
      All lanes : Anti-SMYD3 antibody [EPR11106(2)] - N-terminal (ab183498) at 1/1000 dilution

      Lane 1 : Wild-type HAP1 whole cell lysate
      Lane 2 : SMYD3 knockout HAP1 whole cell lysate
      Lane 3 : HeLa whole cell lysate
      Lane 4 : MCF7 whole cell lysate

      Lysates/proteins at 20 µg per lane.

      Predicted band size: 49 kDa



      Lanes 1 - 4: Merged signal (red and green). Green - ab183498 observed at 49 kDa. Red - loading control, ab8245, observed at 37 kDa.

      ab183498 was shown to recognize SMYD3 in wild-type HAP1 cells as signal was lost at the expected MW in SMYD3 knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and SMYD3 knockout samples were subjected to SDS-PAGE. The membrane was blocked with 3% Milk. Ab183498 and ab8245 (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-SMYD3 antibody [EPR11106(2)] - N-terminal (ab183498)
      Western blot - Anti-SMYD3 antibody [EPR11106(2)] - N-terminal (ab183498)
      All lanes : Anti-SMYD3 antibody [EPR11106(2)] - N-terminal (ab183498) at 1/10000 dilution

      Lane 1 : HeLa lysate
      Lane 2 : 293 lysate
      Lane 3 : T47-D 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: 49 kDa

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SMYD3 antibody [EPR11106(2)] - N-terminal (ab183498)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SMYD3 antibody [EPR11106(2)] - N-terminal (ab183498)

      Immunohistochemical analysis of paraffin-embedded Human skeletal muscle labeling SMYD3 at 1/100 dilution and HRP Polymer for Rabbit IgG. Counterstained with Hematoxylin.

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

    • Immunocytochemistry/ Immunofluorescence - Anti-SMYD3 antibody [EPR11106(2)] - N-terminal (ab183498)
      Immunocytochemistry/ Immunofluorescence - Anti-SMYD3 antibody [EPR11106(2)] - N-terminal (ab183498)

      Flow cytometric analysis of HeLa cells fixed in 20 °C acetone labeling SMYD3 at 1/500 dilution and Goat anti rabbit IgG(Alexa Fluor® 555) at 1/200 (red). Counterstained with DAPI (blue)

    • Immunoprecipitation - Anti-SMYD3 antibody [EPR11106(2)] - N-terminal (ab183498)
      Immunoprecipitation - Anti-SMYD3 antibody [EPR11106(2)] - N-terminal (ab183498)

      Immunoprecipitation of HeLa cells labeling SMYD3 with ab183498 at 1/50 dilution and Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/1500.  

    • Flow Cytometry (Intracellular) - Anti-SMYD3 antibody [EPR11106(2)] - N-terminal (ab183498)
      Flow Cytometry (Intracellular) - Anti-SMYD3 antibody [EPR11106(2)] - N-terminal (ab183498)

      Intracellular flow cytometric analysis of 293 cells fixed in 2% paraformaldehyde labeling SMYD3 with ab183498 at 1/110 dilution andGoat anti rabbit IgG (FITC) at 1/150 dilution. Rabbit Monoclonal IgG was used as an isotype control. 

       

       

    • Western blot - Anti-SMYD3 antibody [EPR11106(2)] - N-terminal (ab183498)
      Western blot - Anti-SMYD3 antibody [EPR11106(2)] - N-terminal (ab183498)
      Anti-SMYD3 antibody [EPR11106(2)] - N-terminal (ab183498) at 1/2000 dilution + MCF-7 lysate at 20 µg

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

      Predicted band size: 49 kDa

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SMYD3 antibody [EPR11106(2)] - N-terminal (ab183498)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SMYD3 antibody [EPR11106(2)] - N-terminal (ab183498)

      Immunohistochemical analysis of paraffin-embedded Human testis labeling SMYD3 with ab183498 at 1/100 dilution and  HRP Polymer for Rabbit IgG. Counterstained with Hematoxylin.

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

    • Anti-SMYD3 antibody [EPR11106(2)] - N-terminal (ab183498)
      Anti-SMYD3 antibody [EPR11106(2)] - N-terminal (ab183498)

    References (2)

    Publishing research using ab183498? Please let us know so that we can cite the reference in this datasheet.

    ab183498 has been referenced in 2 publications.

    • Alshiraihi IM  et al. In Silico/In Vitro Hit-to-Lead Methodology Yields SMYD3 Inhibitor That Eliminates Unrestrained Proliferation of Breast Carcinoma Cells. Int J Mol Sci 21:N/A (2020). PubMed: 33333978
    • McNulty DE  et al. A High-Throughput Dose-Response Cellular Thermal Shift Assay for Rapid Screening of Drug Target Engagement in Living Cells, Exemplified Using SMYD3 and IDO1. SLAS Discov 23:34-46 (2018). PubMed: 28957646

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