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Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-KDM4A / JHDM3A / JMJD2A antibody [EPR18466] - BSA and Azide free (ab251084)

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Western blot - Anti-KDM4A / JHDM3A / JMJD2A antibody [EPR18466] - BSA and Azide free (ab251084)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-KDM4A / JHDM3A / JMJD2A antibody [EPR18466] - BSA and Azide free (ab251084)
  • Immunoprecipitation - Anti-KDM4A / JHDM3A / JMJD2A antibody [EPR18466] - BSA and Azide free (ab251084)
  • Western blot - Anti-KDM4A / JHDM3A / JMJD2A antibody [EPR18466] - BSA and Azide free (ab251084)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-KDM4A / JHDM3A / JMJD2A antibody [EPR18466] - BSA and Azide free (ab251084)
  • Western blot - Anti-KDM4A / JHDM3A / JMJD2A antibody [EPR18466] - BSA and Azide free (ab251084)
  • Anti-KDM4A / JHDM3A / JMJD2A antibody [EPR18466] - BSA and Azide free (ab251084)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR18466] to KDM4A / JHDM3A / JMJD2A - BSA and Azide free
  • Suitable for: IP, WB, IHC-P
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

Conjugates logo Related conjugates and formulations

Unconjugated

Overview

  • Product name

    Anti-KDM4A / JHDM3A / JMJD2A antibody [EPR18466] - BSA and Azide free
    See all KDM4A / JHDM3A / JMJD2A primary antibodies

  • Description

    Rabbit monoclonal [EPR18466] to KDM4A / JHDM3A / JMJD2A - BSA and Azide free

  • Host species

    Rabbit

  • Specificity

    IHC application is recommended for human and rat.

  • Tested applications

    Suitable for: IP, WB, IHC-Pmore details

  • Species reactivity

    Reacts with: Mouse, Rat, Human

  • Immunogen

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

  • General notes

    ab251084 is the carrier-free version of ab191433.

    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.

Properties

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab251084 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
IP
Use at an assay dependent concentration.
WB
Use at an assay dependent concentration. Detects a band of approximately 150 kDa (predicted molecular weight: 120 kDa).
IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Notes
IP
Use at an assay dependent concentration.
WB
Use at an assay dependent concentration. Detects a band of approximately 150 kDa (predicted molecular weight: 120 kDa).
IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Target

  • Function

    Histone demethylase that specifically demethylates 'Lys-9' and 'Lys-36' residues of histone H3, thereby playing a central role in histone code. Does not demethylate histone H3 'Lys-4', H3 'Lys-27' nor H4 'Lys-20'. Demethylates trimethylated H3 'Lys-9' and H3 'Lys-36' residue, while it has no activity on mono- and dimethylated residues. Demethylation of Lys residue generates formaldehyde and succinate. Participates in transcriptional repression of ASCL2 and E2F-responsive promoters via the recruitment of histone deacetylases and NCOR1, respectively.
    Isoform 2: Crucial for muscle differentiation, promotes transcriptional activation of the Myog gene by directing the removal of repressive chromatin marks at its promoter. Lacks the N-terminal demethylase domain.

  • Tissue specificity

    Ubiquitous.

  • Sequence similarities

    Belongs to the JHDM3 histone demethylase family.
    Contains 1 C2HC pre-PHD-type zinc finger.
    Contains 1 JmjC domain.
    Contains 1 JmjN domain.
    Contains 2 PHD-type zinc fingers.
    Contains 2 Tudor domains.

  • Domain

    The 2 Tudor domains recognize and bind methylated histone H3 'Lys-4' residue (H3K4me). Double Tudor domain has an interdigitated structure and the unusual fold is required for its ability to bind methylated histone tails. Trimethylated H3 'Lys-4' (H3K4me3) is bound in a cage of 3 aromatic residues, 2 of which are from the Tudor domain 2, while the binding specificity is determined by side-chain interactions involving residues from the Tudor domain 1. The Tudor domains are also able to bind trimethylated histone H3 'Lys-9' (H3K9me3), di- and trimethylated H4 'Lys-20' (H4K20me2 and H4K20me3). Has high affinity for H4K20me2, blocking recruitment of proteins such as TP53BP1.

  • Post-translational
    modifications

    Ubiquitinated by RNF8 and RNF168 following DNA damage, leading to its degradation. Degradation promotes accessibility of H4K20me2 mark for DNA repair protein TP53BP1, which is then recruited.

  • Cellular localization

    Nucleus.
  • Information by UniProt

  • Database links

    • Alternative names

      • JHDM3A antibody
      • JmjC domain containing histone demethylation protein 3A antibody
      • JmjC domain-containing histone demethylation protein 3A antibody
      • JMJD2 antibody
      • JMJD2A antibody
      • jumonji C domain containing histone demethylase 3A antibody
      • Jumonji domain containing 2 antibody
      • Jumonji domain containing 2A antibody
      • Jumonji domain containing protein 2A antibody
      • Jumonji domain-containing protein 2A antibody
      • KDM4A antibody
      • KDM4A_HUMAN antibody
      • KIAA0677 antibody
      • Lysine (K) specific demethylase 4A antibody
      • Lysine-specific demethylase 4A antibody
      • TDRD14A antibody
      • Tudor domain containing 14A antibody
      see all

    Images

    • Western blot - Anti-KDM4A / JHDM3A / JMJD2A antibody [EPR18466] - BSA and Azide free (ab251084)
      Western blot - Anti-KDM4A / JHDM3A / JMJD2A antibody [EPR18466] - BSA and Azide free (ab251084)

      This data was developed using ab191433, the same antibody clone in a different buffer formulation.

      Lane 1: Wild-type HAP1 cell lysate (20 µg)

      Lane 2: DM4A / JHDM3A / JMJD2A knockout HAP1 cell lysate (20 µg) 

      Lane 3: 2OS cell lysate (20 µg)

      Lane 4: C12 cell lysate (20 µg)

      Lanes 1 - 4: erged signal (red and green). Green - ab191433 observed at 110 kDa. Red - loading control, ab8245, observed at 37 kDa.

      ab191433 was shown to specifically react with KDM4A / JHDM3A / JMJD2A when KDM4A / JHDM3A / JMJD2A knockout samples were used. Wild-type and KDM4A / JHDM3A / JMJD2A knockout samples were subjected to SDS-PAGE. ab191433 and ab8245 (loading control to GAPDH) were diluted 1/500 and 1/10000 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.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-KDM4A / JHDM3A / JMJD2A antibody [EPR18466] - BSA and Azide free (ab251084)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-KDM4A / JHDM3A / JMJD2A antibody [EPR18466] - BSA and Azide free (ab251084)

      This data was developed using ab191433, the same antibody clone in a different buffer formulation.

      Immunohistochemical analysis of paraffin-embedded human colon tissue labeling KDM4A / JHDM3A / JMJD2A with ab191433 at 1/250 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Nuclear staining in epithelial cells of human colon 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-KDM4A / JHDM3A / JMJD2A antibody [EPR18466] - BSA and Azide free (ab251084)
      Immunoprecipitation - Anti-KDM4A / JHDM3A / JMJD2A antibody [EPR18466] - BSA and Azide free (ab251084)

      This data was developed using ab191433, the same antibody clone in a different buffer formulation.

      KDM4A / JHDM3A / JMJD2A was immunoprecipitated from 1mg of HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate with ab191433 at 1/80 dilution. Western blot was performed from the immunoprecipitate using ab191433 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution. Lane 1: HeLa whole cell lysate 10µg (Input). Lane 2: ab191433 IP in HeLa whole cell lysate. Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab191433 in HeLa whole cell lysate. Blocking and dilution buffer and concentration: 5% NFDM/TBST. Exposure time: 3 seconds.
    • Western blot - Anti-KDM4A / JHDM3A / JMJD2A antibody [EPR18466] - BSA and Azide free (ab251084)
      Western blot - Anti-KDM4A / JHDM3A / JMJD2A antibody [EPR18466] - BSA and Azide free (ab251084)
      Lanes 1-3 : Anti-KDM4A / JHDM3A / JMJD2A antibody [EPR18466] (ab191433) at 1/5000 dilution
      Lane 4 : Anti-MTHFD2 antibody [7] (ab191443) at 1/5000 dilution

      Lane 1 : Human fetal kidney at 10 µg
      Lane 2 : HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate at 20 µg
      Lane 3 : LNCaP (Human prostate cancer cell line) whole cell lysate at 20 µg
      Lane 4 : U-2 OS (Human bone osteosarcoma epithelial cell line) whole call lysate at 20 µg

      Secondary
      Lane 1 : Goat Anti-Rabbit IgG Peroxidase Conjugate, specific to the non-reduced form of IgG at 1/100000 dilution
      Lanes 2-3 : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
      Lane 4 : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/5000 dilution

      Developed using the ECL technique.

      Predicted band size: 120 kDa
      Observed band size: 150 kDa why is the actual band size different from the predicted?



      This data was developed using ab191433, the same antibody clone in a different buffer formulation.

      Blocking and dilution buffer: 5% NFDM/TBST.

      Exposure times: Lanes 1, 3-4: 1 minute; Lane 2: 3 minutes.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-KDM4A / JHDM3A / JMJD2A antibody [EPR18466] - BSA and Azide free (ab251084)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-KDM4A / JHDM3A / JMJD2A antibody [EPR18466] - BSA and Azide free (ab251084)

      This data was developed using ab191433, the same antibody clone in a different buffer formulation.

      Immunohistochemical analysis of paraffin-embedded rat colon tissue labeling KDM4A / JHDM3A / JMJD2A with ab191433 at 1/250 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Nuclear staining in epithelial cells of rat colon 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.
    • Western blot - Anti-KDM4A / JHDM3A / JMJD2A antibody [EPR18466] - BSA and Azide free (ab251084)
      Western blot - Anti-KDM4A / JHDM3A / JMJD2A antibody [EPR18466] - BSA and Azide free (ab251084)
      All lanes : Anti-KDM4A / JHDM3A / JMJD2A antibody [EPR18466] (ab191433) at 1/5000 dilution

      Lane 1 : PC-12 (Rat adrenal gland pheochromocytoma cell line) whole cell lysate
      Lane 2 : NIH/3T3 (Mouse embryonic fibroblast cell line) whole cell lysate

      Lysates/proteins at 10 µg per lane.

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

      Developed using the ECL technique.

      Predicted band size: 120 kDa
      Observed band size: 150 kDa why is the actual band size different from the predicted?


      Exposure time: 30 seconds


      This data was developed using ab191433, the same antibody clone in a different buffer formulation.

      Blocking and dilution buffer: 5% NFDM/TBST.

    • Anti-KDM4A / JHDM3A / JMJD2A antibody [EPR18466] - BSA and Azide free (ab251084)
      Anti-KDM4A / JHDM3A / JMJD2A antibody [EPR18466] - BSA and Azide free (ab251084)

    Protocols

    To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

    Click here to view the general protocols

    References (0)

    Publishing research using ab251084? Please let us know so that we can cite the reference in this datasheet.

    ab251084 has not yet been referenced specifically in any publications.

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