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Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-EHMT2/G9A antibody [EPR18894] - BSA and Azide free (ab240289)

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Western blot - Anti-EHMT2/G9A antibody [EPR18894] - BSA and Azide free (ab240289)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-EHMT2/G9A antibody [EPR18894] - BSA and Azide free (ab240289)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-EHMT2/G9A antibody [EPR18894] - BSA and Azide free (ab240289)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-EHMT2/G9A antibody [EPR18894] - BSA and Azide free (ab240289)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-EHMT2/G9A antibody [EPR18894] - BSA and Azide free (ab240289)
  • Immunocytochemistry/ Immunofluorescence - Anti-EHMT2/G9A antibody [EPR18894] - BSA and Azide free (ab240289)
  • Immunocytochemistry/ Immunofluorescence - Anti-EHMT2/G9A antibody [EPR18894] - BSA and Azide free (ab240289)
  • Flow Cytometry (Intracellular) - Anti-EHMT2/G9A antibody [EPR18894] - BSA and Azide free (ab240289)
  • Immunoprecipitation - Anti-EHMT2/G9A antibody [EPR18894] - BSA and Azide free (ab240289)
  • Anti-EHMT2/G9A antibody [EPR18894] - BSA and Azide free (ab240289)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR18894] to EHMT2/G9A - BSA and Azide free
  • Suitable for: WB, Flow Cyt (Intra), ICC/IF, IP, IHC-P
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

Conjugates logo Related conjugates and formulations

Alexa Fluor® 488 Unconjugated

Overview

  • Product name

    Anti-EHMT2/G9A antibody [EPR18894] - BSA and Azide free
    See all EHMT2/G9A primary antibodies

  • Description

    Rabbit monoclonal [EPR18894] to EHMT2/G9A - BSA and Azide free

  • Host species

    Rabbit

  • Tested applications

    Suitable for: WB, Flow Cyt (Intra), ICC/IF, IP, IHC-Pmore details

  • Species reactivity

    Reacts with: Mouse, Rat, Human

  • Immunogen

    Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • WB: HeLa and HEK-293 cell lysates. IHC-P: Human colon and gastric adenocarcinoma tissues; Mouse liver tissue; Rat kidney tissue. ICC/IF: HeLa and MCF7 cells. Flow Cyt (intra): HeLa cells. IP: HeLa cells.

  • General notes

    ab240289 is the carrier-free version of ab185050.

    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 ab240289 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
Use at an assay dependent concentration. Detects a band of approximately 170, 160 kDa (predicted molecular weight: 132 kDa).
Flow Cyt (Intra)
Use at an assay dependent concentration.
ICC/IF
Use at an assay dependent concentration.
IP
Use at an assay dependent concentration.
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
WB
Use at an assay dependent concentration. Detects a band of approximately 170, 160 kDa (predicted molecular weight: 132 kDa).
Flow Cyt (Intra)
Use at an assay dependent concentration.
ICC/IF
Use at an assay dependent concentration.
IP
Use at an assay dependent concentration.
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 methyltransferase that specifically mono- and dimethylates 'Lys-9' of histone H3 (H3K9me1 and H3K9me2, respectively) in euchromatin. H3K9me represents a specific tag for epigenetic transcriptional repression by recruiting HP1 proteins to methylated histones. Also mediates monomethylation of 'Lys-56' of histone H3 (H3K56me1) in G1 phase, leading to promote interaction between histone H3 and PCNA and regulating DNA replication. Also weakly methylates 'Lys-27' of histone H3 (H3K27me). Also required for DNA methylation, the histone methyltransferase activity is not required for DNA methylation, suggesting that these 2 activities function independently. Probably targeted to histone H3 by different DNA-binding proteins like E2F6, MGA, MAX and/or DP1. May also methylate histone H1. In addition to the histone methyltransferase activity, also methylates non-histone proteins: mediates dimethylation of 'Lys-373' of p53/TP53. Also methylates CDYL, WIZ, ACIN1, DNMT1, HDAC1, ERCC6, KLF12 and itself.

  • Tissue specificity

    Expressed in all tissues examined, with high levels in fetal liver, thymus, lymph node, spleen and peripheral blood leukocytes and lower level in bone marrow.

  • Sequence similarities

    Belongs to the class V-like SAM-binding methyltransferase superfamily. Histone-lysine methyltransferase family. Suvar3-9 subfamily.
    Contains 7 ANK repeats.
    Contains 1 post-SET domain.
    Contains 1 pre-SET domain.
    Contains 1 SET domain.

  • Domain

    The SET domain mediates interaction with WIZ.
    The ANK repeats bind H3K9me1 and H3K9me2.

  • Post-translational
    modifications

    Methylated at Lys-185; automethylated.

  • Cellular localization

    Nucleus. Chromosome. Associates with euchromatic regions. Does not associate with heterochromatin.
  • Information by UniProt

  • Database links

    • Alternative names

      • Ankyrin repeat containing protein antibody
      • Bat 8 antibody
      • Bat8 antibody
      • C6orf30 antibody
      • DKFZp686H08213 antibody
      • EHMT 2 antibody
      • Ehmt2 antibody
      • EHMT2_HUMAN antibody
      • Euchromatic histone lysine methyltransferase 2 antibody
      • Euchromatic histone lysine N methyltransferase 2 antibody
      • Euchromatic histone-lysine N-methyltransferase 2 antibody
      • FLJ35547 antibody
      • G 9a antibody
      • G9 a antibody
      • G9a protein antibody
      • G9A antibody
      • G9A histone methyltransferase antibody
      • GAT8 antibody
      • H3 K9 HMTase 3 antibody
      • H3-K9-HMTase 3 antibody
      • Histone H3 K9 methyltransferase 3 antibody
      • Histone H3 K9 methyltransferase3 antibody
      • Histone H3-K9 methyltransferase 3 antibody
      • Histone lysine N methyltransferase antibody
      • Histone lysine N methyltransferase EHMT2 antibody
      • Histone lysine N methyltransferase, H3 lysine 9 specific 3 antibody
      • Histone lysine N methyltransferase, H3 lysine 9 specific3 antibody
      • Histone-lysine N-methyltransferase EHMT2 antibody
      • HLA B associated transcript 8 antibody
      • HLA-B-associated transcript 8 antibody
      • KMT 1C antibody
      • KMT1 C antibody
      • Lysine N methyltransferase 1C antibody
      • Lysine N-methyltransferase 1C antibody
      • NG 36 antibody
      • NG36 antibody
      • Protein G9a antibody
      see all

    Images

    • Western blot - Anti-EHMT2/G9A antibody [EPR18894] - BSA and Azide free (ab240289)
      Western blot - Anti-EHMT2/G9A antibody [EPR18894] - BSA and Azide free (ab240289)
      All lanes : Anti-EHMT2/G9A antibody [EPR18894] (ab185050) at 1/1000 dilution

      Lane 1 : Wild-type HeLa cell lysate
      Lane 2 : EHMT2/G9A knockout HeLa cell lysate
      Lane 3 : HEK-293 cell lysate
      Lane 4 : HepG2 cell lysate

      Lysates/proteins at 20 µg per lane.

      Performed under reducing conditions.

      Predicted band size: 132 kDa



      This data was developed using the same antibody clone in a different buffer formulation (ab185050).

      Lanes 1-4: Merged signal (red and green). Green - ab185050 observed at 160 kDa. Red - loading control ab8245 observed at 37 kDa.

       ab185050 Anti-EHMT2/G9A antibody [EPR18894] was shown to specifically react with EHMT2/G9A in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265149 (knockout cell lysate ab257080) was used. Wild-type and EHMT2/G9A knockout samples were subjected to SDS-PAGE. ab185050 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. 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 in 20000 dilution for 1 hour at room temperature before imaging.

       

       

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-EHMT2/G9A antibody [EPR18894] - BSA and Azide free (ab240289)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-EHMT2/G9A antibody [EPR18894] - BSA and Azide free (ab240289)

      Immunohistochemical analysis of paraffin-embedded Human colon tissue labeling EHMT2/G9A with ab185050 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

      Nucleus staining on epithelial cells of the normal 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.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab185050).

      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-EHMT2/G9A antibody [EPR18894] - BSA and Azide free (ab240289)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-EHMT2/G9A antibody [EPR18894] - BSA and Azide free (ab240289)

      Immunohistochemical analysis of paraffin-embedded Human gastric adenocarcinoma tissue labeling EHMT2/G9A with ab185050 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

      Nucleus staining on tumor cells of the gastric adenocarcinoma 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.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab185050).

      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-EHMT2/G9A antibody [EPR18894] - BSA and Azide free (ab240289)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-EHMT2/G9A antibody [EPR18894] - BSA and Azide free (ab240289)

      Immunohistochemical analysis of paraffin-embedded Mouse liver tissue labeling EHMT2/G9A with ab185050 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

      Nucleus staining on hepatocytes of the mouse liver 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.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab185050).

      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-EHMT2/G9A antibody [EPR18894] - BSA and Azide free (ab240289)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-EHMT2/G9A antibody [EPR18894] - BSA and Azide free (ab240289)

      Immunohistochemical analysis of paraffin-embedded Rat kidney tissue labeling EHMT2/G9A with ab185050 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

      Nucleus staining on rat kidney 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.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab185050).

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

    • Immunocytochemistry/ Immunofluorescence - Anti-EHMT2/G9A antibody [EPR18894] - BSA and Azide free (ab240289)
      Immunocytochemistry/ Immunofluorescence - Anti-EHMT2/G9A antibody [EPR18894] - BSA and Azide free (ab240289)

      Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (Human epithelial cells from cervix adenocarcinoma) cells labeling EHMT2/G9A with ab185050 at 1/1000 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green).

      Confocal image showing nuclear staining on HeLa cell line.

      The nuclear counterstain is DAPI (blue).

      Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/1000 dilution (red).

      The negative controls are as follows:-

      -ve control 1: ab185050 at 1/1000 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/1000 dilution.

      -ve control 2: ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/1000 dilution.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab185050).

    • Immunocytochemistry/ Immunofluorescence - Anti-EHMT2/G9A antibody [EPR18894] - BSA and Azide free (ab240289)
      Immunocytochemistry/ Immunofluorescence - Anti-EHMT2/G9A antibody [EPR18894] - BSA and Azide free (ab240289)

      Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized MCF7 (Human breast adenocarcinoma cell line) cells labeling EHMT2/G9A with ab185050 at 1/100 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green).

      Confocal image showing no staining on MCF7 cell line, as MCF7 cells have a very low level expression of EHMT2/G9A. 

      The nuclear counterstain is DAPI (blue).

      Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/1000 dilution (red).

      The negative controls are as follows:-

      -ve control 1: ab185050 at 1/70 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/1000 dilution.

      -ve control 2: ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/1000 dilution.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab185050).

    • Flow Cytometry (Intracellular) - Anti-EHMT2/G9A antibody [EPR18894] - BSA and Azide free (ab240289)
      Flow Cytometry (Intracellular) - Anti-EHMT2/G9A antibody [EPR18894] - BSA and Azide free (ab240289)

      Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed HeLa (Human epithelial cells from cervix adenocarcinoma) cells labeling  EHMT2/G9A with ab185050 at 1/150 dilution (red) compared with a rabbit monoclonal IgG isotype control (ab172730; black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody; blue). Goat anti rabbit IgG (FITC) at 1/150 dilution was used as the secondary antibody. 

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab185050). 

       

       

    • Immunoprecipitation - Anti-EHMT2/G9A antibody [EPR18894] - BSA and Azide free (ab240289)
      Immunoprecipitation - Anti-EHMT2/G9A antibody [EPR18894] - BSA and Azide free (ab240289)

      EHMT2/G9A was immunoprecipitated from 1mg of HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell lysate with ab185050 at 1/50 dilution.

      Western blot was performed from the immunoprecipitate using ab185050 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 10ug (Input).

      Lane 2: ab185050 IP in HeLa whole cell lysate.

      Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab185050 in HeLa whole cell lysate.

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

      Exposure time: 30 seconds.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab185050).

    • Anti-EHMT2/G9A antibody [EPR18894] - BSA and Azide free (ab240289)
      Anti-EHMT2/G9A antibody [EPR18894] - BSA and Azide free (ab240289)

    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 ab240289? Please let us know so that we can cite the reference in this datasheet.

    ab240289 has not yet been referenced specifically in any publications.

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