Validated using a knockout cell line
Recombinant
RabMAb

Recombinant Anti-KMT6 / EZH2 antibody [EPR9307(2)] - BSA and Azide free (ab231165)

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Overview

  • Product name

    Anti-KMT6 / EZH2 antibody [EPR9307(2)] - BSA and Azide free
    See all KMT6 / EZH2 primary antibodies

  • Description

    Rabbit monoclonal [EPR9307(2)] to KMT6 / EZH2 - BSA and Azide free

  • Host species

    Rabbit

  • Tested applications

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

  • Species reactivity

    Reacts with: Mouse, Rat, Human

  • Immunogen

    Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) within Human KMT6/ EZH2 aa 1-100 (N terminal). The exact sequence is proprietary.
    Database link: Q15910

  • Positive control

    • Human breast carcinoma tissue; HeLa cells.

  • General notes

    Ab231165 is the carrier-free version of ab191080. This format is designed for use in antibody labeling, including fluorochromes, metal isotopes, oligonucleotides, enzymes.

     

    Our carrier-free formats are supplied in a buffer free of BSA, sodium azide and glycerol for higher conjugation efficiency.

    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.

    ab231165 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.

    Maxpar® is a trademark of Fluidigm Canada Inc.

Properties

Applications

Our Abpromise guarantee covers the use of ab231165 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
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.
ICC/IF Use at an assay dependent concentration.
WB Use at an assay dependent concentration. Detects a band of approximately 93 kDa (predicted molecular weight: 85 kDa).
Flow Cyt Use at an assay dependent concentration.

ab199376 - Rabbit monoclonal IgG (Low endotoxin, Azide free), is suitable for use as an isotype control with this antibody.

 

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

    see all

  • 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

  • Western blot - Anti-KMT6 / EZH2 antibody [EPR9307(2)] - BSA and Azide free (ab231165)
    Western blot - Anti-KMT6 / EZH2 antibody [EPR9307(2)] - BSA and Azide free (ab231165)

    This WB data was generated using the same anti-KMT6/EZH2 antibody clone, EPR9307(2), in a different buffer formulation (cat# ab191080).

    Lane 1: Wild-type HAP1 cell lysate (20 µg)
    Lane 2: KMT6 / EZH2 knockout HAP1 cell lysate (20 µg) 
    Lane 3: HEK293 cell lysate (20 µg)
    Lane 4: Ms testis tissue lysate (20 µg)
    Lanes 1 - 4: Merged signal (red and green). Green - ab191080 observed at 93 kDa. Red - loading control, ab8245, observed at 37 kDa.

    ab191080 was shown to recognize KMT6 / EZH2 when KMT6 / EZH2 knockout samples were used, along with additional cross-reactive bands. Wild-type and KMT6 / EZH2 knockout samples were subjected to SDS-PAGE. ab191080 and ab8245 (loading control to GAPDH) were diluted at 1/500 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 h at room temperature before imaging.

  • Flow Cytometry - Anti-KMT6 / EZH2 antibody [EPR9307(2)] - BSA and Azide free (ab231165)
    Flow Cytometry - Anti-KMT6 / EZH2 antibody [EPR9307(2)] - BSA and Azide free (ab231165)

    ab191080 staining KMT6 / EZH2 in the human cell line Jurkat (human acute T cell leukemia) by flow cytometry. Cells were fixed with 4% paraformaldehyde and the sample was incubated with the primary antibody at a dilution of 1/90. A goat anti rabbit IgG (Alexa Fluor® 488) at a dilution of 1/2000 was used as the secondary antibody.

    Isoytype control: Rabbit monoclonal IgG (Black)

    Unlabelled control: Cell without incubation with primary antibody and secondary antibody (Blue)

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab191080).

  • Immunocytochemistry/ Immunofluorescence - Anti-KMT6 / EZH2 antibody [EPR9307(2)] - BSA and Azide free (ab231165)
    Immunocytochemistry/ Immunofluorescence - Anti-KMT6 / EZH2 antibody [EPR9307(2)] - BSA and Azide free (ab231165)

    Immunofluorescent analysis of HeLa cells (4% Paraformaldehyde-fixed; 0.1% tritonX-100-permeabilized) labeling KMT6 / EZH2 with ab191080 at 1/250 dilution followed by Goat anti rabbit IgG (AlexaFluor® 488) secondary at 1/200 dilution and counter-stained with DAPI (blue). 

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab191080).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-KMT6 / EZH2 antibody [EPR9307(2)] - BSA and Azide free (ab231165)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-KMT6 / EZH2 antibody [EPR9307(2)] - BSA and Azide free (ab231165)

    This IHC data was generated using the same anti-KMT6/EZH2 antibody clone, EPR9307(2), in a different buffer formulation (cat# ab191080).

    Immunohistochemical analysis of paraffin-embedded Human breast carcinoma tissue labeling KMT6 / EZH2 with ab191080 at 1/250 dilution followed by pre-diluted HRP Polymer for Rabbit IgG secondary antibody and counter-stained with Hematoxylin (inset: negative control).

    Heat mediated antigen retrieval was performed with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

References

ab231165 has not yet been referenced specifically in any publications.

Publishing research using ab231165? Please let us know so that we can cite the reference in this datasheet.

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