Overview

  • Product name
    Anti-KMT6 / EZH2 antibody [EPR20108]
    See all KMT6 / EZH2 primary antibodies
  • Description
    Rabbit monoclonal [EPR20108] to KMT6 / EZH2
  • Host species
    Rabbit
  • Tested applications
    Suitable for: WB, IPmore details
  • Species reactivity
    Reacts with: Human
  • Immunogen

    Synthetic peptide within Human KMT6/ EZH2 aa 350-450. The exact sequence is proprietary.
    Database link: Q15910

  • Positive control
    • WB: MCF7, HeLa, U-2 OS and HEK-293 whole cell lysates; Wild-type HAP1 cell lysate. IP: HeLa whole cell lysate.
  • General notes

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab191250 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 1/1000. Detects a band of approximately 90 kDa (predicted molecular weight: 85 kDa).
IP 1/30.

Target

  • Function
    Polycomb group (PcG) protein. Catalytic subunit of the PRC2/EED-EZH2 complex, which methylates 'Lys-9' and 'Lys-27' of histone H3, leading to transcriptional repression of the affected target gene. Able to mono-, di- and trimethylate 'Lys-27' of histone H3 to form H3K27me1, H3K27me2 and H3K27me3, respectively. Compared to EZH2-containing complexes, it is more abundant in embryonic stem cells and plays a major role in forming H3K27me3, which is required for embryonic stem cell identity and proper differentiation. The PRC2/EED-EZH2 complex may also serve as a recruiting platform for DNA methyltransferases, thereby linking two epigenetic repression systems. Genes repressed by the PRC2/EED-EZH2 complex include HOXC8, HOXA9, MYT1, CDKN2A and retinoic acid target genes.
  • Tissue specificity
    Expressed in many tissues. Overexpressed in numerous tumor types including carcinomas of the breast, colon, larynx, lymphoma and testis.
  • Sequence similarities
    Belongs to the histone-lysine methyltransferase family. EZ subfamily.
    Contains 1 SET domain.
  • Developmental stage
    Expression decreases during senescence of embryonic fibroblasts (HEFs). Expression peaks at the G1/S phase boundary.
  • Post-translational
    modifications
    Phosphorylated by AKT1. Phosphorylation by AKT1 reduces methyltransferase activity.
  • Cellular localization
    Nucleus.
  • Information by UniProt
  • Database links
  • Alternative names
    • Enhancer of zeste 2 antibody
    • enhancer of zeste 2 polycomb repressive complex 2 subunit antibody
    • Enhancer of zeste homolog 2 (Drosophila) antibody
    • Enhancer of zeste homolog 2 antibody
    • Enhancer of zeste, Drosophila, homolog 2 antibody
    • ENX 1 antibody
    • Enx 1h antibody
    • ENX-1 antibody
    • ENX1 antibody
    • Enx1h antibody
    • EZH 2 antibody
    • EZH1 antibody
    • EZH2 antibody
    • EZH2_HUMAN antibody
    • EZH2b antibody
    • Histone-lysine N-methyltransferase EZH2 antibody
    • KMT 6 antibody
    • KMT6 antibody
    • KMT6A antibody
    • Lysine N-methyltransferase 6 antibody
    • MGC9169 antibody
    • WVS antibody
    • WVS2 antibody
    see all

Images

  • All lanes : Anti-KMT6 / EZH2 antibody [EPR20108] (ab191250) at 1/1000 dilution

    Lane 1 : Wild-type HAP1 cell lysate
    Lane 2 : KMT6 / EZH2 knockout HAP1 cell lysate
    Lane 3 : HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate
    Lane 4 : A549 (Human lung carcinoma cell line) whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Predicted band size: 85 kDa
    Observed band size: 90 kDa
    why is the actual band size different from the predicted?



    Merged signal (red and green). Green - ab191250 observed at 93 kDa. Red - loading control, ab8245, observed at 37 kDa.

    ab191250 was shown to specifically recognize KMT6/EZH2 in wild-type HAP1 cells along with additional cross-reactive bands. No band was observed when KMT6/EZH2 knockout samples were examined. Wild-type and KMT6/EZH2 knockout samples were subjected to SDS-PAGE. ab191250 and ab8245 (loading control to GAPDH) were diluted at 1/1000 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 1/10,000 dilution for 1 hour at room temperature before imaging.

  • All lanes : Anti-KMT6 / EZH2 antibody [EPR20108] (ab191250) at 1/5000 dilution

    Lane 1 : MCF7 (Human breast adenocarcinoma cell line) whole cell lysate
    Lane 2 : HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate

    Lysates/proteins at 20 µg per lane.

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

    Predicted band size: 85 kDa
    Observed band size: 90 kDa why is the actual band size different from the predicted?


    Exposure time: 30 seconds


    Blocking/Dilution buffer: 5% NFDM/TBST.

  • Anti-KMT6 / EZH2 antibody [EPR20108] (ab191250) at 1/1000 dilution + U-2 OS (Human bone osteosarcoma epithelial cell line) whole cell lysate at 20 µg

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

    Predicted band size: 85 kDa
    Observed band size: 90 kDa why is the actual band size different from the predicted?


    Exposure time: 30 seconds


    Blocking/Dilution buffer: 5% NFDM/TBST.

  • KMT6 / EZH2 was immunoprecipitated from 0.35 mg of HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate with ab191250 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab191250 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 10 μg (Input).

    Lane 2: ab191250 IP in HeLa whole cell lysate.

    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab191250 in HeLa whole cell lysate.

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

    Exposure time: 3 minutes.

References

This product has been referenced in:
  • Xu K  et al. LncRNA FOXD2-AS1 plays an oncogenic role in hepatocellular carcinoma through epigenetically silencing CDKN1B(p27) via EZH2. Exp Cell Res 380:198-204 (2019). Read more (PubMed: 31004581) »
  • Li D  et al. Armadillo repeat containing 12 promotes neuroblastoma progression through interaction with retinoblastoma binding protein 4. Nat Commun 9:2829 (2018). Read more (PubMed: 30026490) »
See all 3 Publications for this product

Customer reviews and Q&As

Application
ChIP
Sample
Human Cell lysate - nuclear (Human B cells)
Negative control
Isotype IgG
Specification
Human B cells
Detection step
Real-time PCR
Type
Cross-linking (X-ChIP)
Duration of cross-linking step: 5 minute(s) and 0 second(s)
Specification of the cross-linking agent: Formaldehyde
Positive control
Gene specific Promoter primers

Abcam user community

Verified customer

Submitted Jul 05 2017

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