Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-KDM3A / JHDM2A antibody [EPR18875] (ab191389)

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Western blot - Anti-KDM3A / JHDM2A antibody [EPR18875] (ab191389)
  • Western blot - Anti-KDM3A / JHDM2A antibody [EPR18875] (ab191389)
  • Western blot - Anti-KDM3A / JHDM2A antibody [EPR18875] (ab191389)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-KDM3A / JHDM2A antibody [EPR18875] (ab191389)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-KDM3A / JHDM2A antibody [EPR18875] (ab191389)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-KDM3A / JHDM2A antibody [EPR18875] (ab191389)
  • Immunoprecipitation - Anti-KDM3A / JHDM2A antibody [EPR18875] (ab191389)
  • Anti-KDM3A / JHDM2A antibody [EPR18875] (ab191389)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR18875] to KDM3A / JHDM2A
  • Suitable for: WB, IHC-P, IP
  • Knockout validated
  • Reacts with: Human

Conjugates logo Related conjugates and formulations

Carrier Free

Overview

  • Product name

    Anti-KDM3A / JHDM2A antibody [EPR18875]
    See all KDM3A / JHDM2A primary antibodies

  • Description

    Rabbit monoclonal [EPR18875] to KDM3A / JHDM2A

  • Host species

    Rabbit

  • Tested applications

    Suitable for: WB, IHC-P, IPmore details

  • Species reactivity

    Reacts with: Human

  • Immunogen

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

  • Positive control

    • WB: Untreated HEK-293 whole cell lysate; HEK-293, HeLa and Jurkat treated with 100 µM CoCl2 for 24 hours whole cell lysates; Ramos whole cell lysate. IHC-P: Human testis, bladder cancer and lung cancer tissues. IP: Ramos whole cell lysate.

  • 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 ab191389 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)
1/1000. Detects a band of approximately 147, 100, 75 kDa (predicted molecular weight: 147 kDa).
IHC-P
1/1000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
IP
1/30.
Notes
WB
1/1000. Detects a band of approximately 147, 100, 75 kDa (predicted molecular weight: 147 kDa).
IHC-P
1/1000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
IP
1/30.

Target

  • Function

    Histone demethylase that specifically demethylates 'Lys-9' of histone H3, thereby playing a central role in histone code. Preferentially demethylates mono- and dimethylated H3 'Lys-9' residue, with a preference for dimethylated residue, while it has weak or no activity on trimethylated H3 'Lys-9'. Demethylation of Lys residue generates formaldehyde and succinate. Involved in hormone-dependent transcriptional activation, by participating in recruitment to androgen-receptor target genes, resulting in H3 'Lys-9' demethylation and transcriptional activation. Involved in spermatogenesis by regulating expression of target genes such as PRM1 and TMP1 which are required for packaging and condensation of sperm chromatin. Involved in obesity resistance through regulation of metabolic genes such as PPARA and UCP1.

  • Sequence similarities

    Belongs to the JHDM2 histone demethylase family.
    Contains 1 JmjC domain.

  • Domain

    The JmjC domain and the C6-type zinc-finger are required for the demethylation activity.
    Leu-Xaa-Xaa-Leu-Leu (LXXLL) motifs are known to mediate the association with nuclear receptors.

  • Cellular localization

    Cytoplasm. Nucleus. Nuclear in round spermatids. When spermatids start to elongate, localizes to the cytoplasm where it forms distinct foci which disappear in mature spermatozoa.
  • Information by UniProt

  • Database links

    • Alternative names

      • DKFZp686A24246 antibody
      • DKFZp686P07111 antibody
      • JHDM2A antibody
      • JHMD2A antibody
      • JmjC domain-containing histone demethylation protein 2A antibody
      • JMJD1 antibody
      • JMJD1A antibody
      • Jumonji C domain containing histone demethylase 2A antibody
      • Jumonji domain containing 1 antibody
      • Jumonji domain containing 1A antibody
      • Jumonji domain containing protein 1A antibody
      • Jumonji domain-containing protein 1A antibody
      • Kdm3a antibody
      • KDM3A_HUMAN antibody
      • KIAA0742 antibody
      • Lysine (K) specific demethylase 3A antibody
      • Lysine-specific demethylase 3A antibody
      • Testis specific protein A antibody
      • TSGA antibody
      see all

    Images

    • Western blot - Anti-KDM3A / JHDM2A antibody [EPR18875] (ab191389)
      Western blot - Anti-KDM3A / JHDM2A antibody [EPR18875] (ab191389)

      Lane 1: Wild type HAP1 whole cell lysate (40 µg)
      Lane 2: KDM3A knockout  HAP1 whole cell lysate (40 µg)
      Lane 3: HeLa whole cell lysate (40 µg)
      Lane 4: Hek293 whole cell lysate (40 µg)
      Lanes 1 - 4: Merged signal (red and green). Green - ab191389 observed at 150 kDa. Red - loading control, ab18058, observed at 130 kDa.

      ab191389 was shown to specifically react with KDM3A when KDM3A knockout samples were used. Wild-type and KDM3A knockout samples were subjected to SDS-PAGE.  Ab191389 and ab18058 (Mouse anti Vinculin loading control) were incubated overnight at 4°C at 500 dilution and 1/10000 dilution respectively. Blots were developed with 800CW Goat anti Rabbit and 680CW Goat anti Mouse secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.

       

    • Western blot - Anti-KDM3A / JHDM2A antibody [EPR18875] (ab191389)
      Western blot - Anti-KDM3A / JHDM2A antibody [EPR18875] (ab191389)
      All lanes : Anti-KDM3A / JHDM2A antibody [EPR18875] (ab191389) at 1/1000 dilution

      Lane 1 : Untreated HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate
      Lane 2 : HEK-293 (Human epithelial cell line from embryonic kidney) treated with 100 µM CoCl2 for 24 hours whole cell lysate
      Lane 3 : Untreated HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate
      Lane 4 : HeLa (Human epithelial cell line from cervix adenocarcinoma) treated with 100 µM CoCl2 for 24 hours whole cell lysate
      Lane 5 : Untreated Jurkat (Human T cell leukemia cell line from peripheral blood) whole cell lysate
      Lane 6 : Jurkat (Human T cell leukemia cell line from peripheral blood) treated with 100 µM CoCl2 for 24 hours whole cell lysate

      Lysates/proteins at 20 µg per lane.

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

      Predicted band size: 147 kDa
      Observed band size: 100,147,75 kDa why is the actual band size different from the predicted?



      Blocking/Dilution buffer: 5% NFDM/TBST.

      Exposure time: Lane 1 and 2: 15 seconds; Lane 3 and 4: 30 seconds; Lane 5 and 6: 5 seconds.

       

    • Western blot - Anti-KDM3A / JHDM2A antibody [EPR18875] (ab191389)
      Western blot - Anti-KDM3A / JHDM2A antibody [EPR18875] (ab191389)
      Anti-KDM3A / JHDM2A antibody [EPR18875] (ab191389) at 1/1000 dilution + Ramos (Human Burkitt's lymphoma cell line) whole cell lysate

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

      Predicted band size: 147 kDa
      Observed band size: 147 kDa


      Exposure time: 3 minutes


      Blocking/Dilution buffer: 5% NFDM/TBST.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-KDM3A / JHDM2A antibody [EPR18875] (ab191389)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-KDM3A / JHDM2A antibody [EPR18875] (ab191389)

      Immunohistochemical analysis of paraffin-embedded human testis tissue labeling KDM3A / JHDM2A with ab191389 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

      Nuclear staining on spermatogenic cells of human testis is observed.

      Counter stained with Hematoxylin.

      Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

      Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-KDM3A / JHDM2A antibody [EPR18875] (ab191389)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-KDM3A / JHDM2A antibody [EPR18875] (ab191389)

      Immunohistochemical analysis of paraffin-embedded human bladder cancer tissue labeling KDM3A / JHDM2A with ab191389 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

      Weak nuclear staining on cancer cells of human bladder cancer is observed.

      Counter stained with Hematoxylin.

      Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

      Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-KDM3A / JHDM2A antibody [EPR18875] (ab191389)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-KDM3A / JHDM2A antibody [EPR18875] (ab191389)

      Immunohistochemical analysis of paraffin-embedded human lung cancer tissue labeling KDM3A / JHDM2A with ab191389 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

      Weak nuclear staining on cancer cells of human lung cancer is observed.

      Counter stained with Hematoxylin.

      Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

      Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

    • Immunoprecipitation - Anti-KDM3A / JHDM2A antibody [EPR18875] (ab191389)
      Immunoprecipitation - Anti-KDM3A / JHDM2A antibody [EPR18875] (ab191389)

      KDM3A / JHDM2A was immunoprecipitated from 1mg of Ramos (Human Burkitt's lymphoma cell line) whole cell lysate with ab191389 at 1/30 dilution.

      Western blot was performed from the immunoprecipitate using ab191389 at 1/1000 dilution.

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

      Lane 1: Ramos whole cell lysate 10µg (Input).

      Lane 2: ab191389 IP in Ramos whole cell lysate.

      Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab191389 in Ramos whole cell lysate.

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

      Exposure time: 10 seconds.

    • Anti-KDM3A / JHDM2A antibody [EPR18875] (ab191389)
      Anti-KDM3A / JHDM2A antibody [EPR18875] (ab191389)

    References (1)

    Publishing research using ab191389? Please let us know so that we can cite the reference in this datasheet.

    ab191389 has been referenced in 1 publication.

    • Qian X  et al. KDM3A Senses Oxygen Availability to Regulate PGC-1a-Mediated Mitochondrial Biogenesis. Mol Cell 76:885-895.e7 (2019). PubMed: 31629659

    Customer reviews and Q&As

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    Western blot abreview for Anti-KDM3A / JHDM2A antibody [EPR18875]

     Poor
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Mouse Tissue lysate - whole (E18.5 Liver)
    Gel Running Conditions
    Reduced Denaturing
    Loading amount
    50 µg
    Specification
    E18.5 Liver
    Blocking step
    Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
    Read More

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    Submitted Sep 06 2018

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