Overview

  • Product name
    Anti-EHMT2/G9A antibody [EPR18894]
    See all EHMT2/G9A primary antibodies
  • Description
    Rabbit monoclonal [EPR18894] to EHMT2/G9A
  • Host species
    Rabbit
  • Tested applications
    Suitable for: WB, IHC-P, ICC/IF, Flow Cyt, IPmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Recombinant fragment within Human EHMT2/G9A aa 1-300. The exact sequence is proprietary.
    Database link: Q96KQ7

  • Positive control
    • WB: HEK-293, Jurkat, HepG2, NCCIT, F9, Neuro-2a, LLC1, C6, RAW 264.7, PC-12 and NIH/3T3 whole cell lysates. Human fetal heart and fetal kidney lysates. IHC-P: Human colon, Human gastric adenocarcinoma, mouse liver and rat kidney tissues. ICC/IF: HeLa. Flow Cyt: HeLa cells. 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 ab185050 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 170, 160 kDa (predicted molecular weight: 132 kDa).
IHC-P 1/500. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
ICC/IF 1/1000.
Flow Cyt 1/150.
IP 1/50.

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

  • Lane 1: Wild-type HAP1 cell lysate (20 µg)
    Lane 2: EHMT2/G9A knockout HAP1 cell lysate (20 µg)
    Lane 3: HEK293 cell lysate (20 µg)
    Lane 4: HepG2 cell lysate (20 µg)

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

    ab185050 was shown to recognize EHMT2/G9A when EHMT2/G9A knockout samples were used, along with additional cross-reactive bands. Wild-type and EHMT2/G9A knockout samples were subjected to SDS-PAGE. ab185050 and ab8245 (loading control to GAPDH) were diluted 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 h at room temperature before imaging.

  • 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.

  • 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.

  • All lanes : Anti-EHMT2/G9A antibody [EPR18894] (ab185050) at 1/1000 dilution

    Lane 1 : HEK-293 (Human epithelial cells from embryonic kidney) whole cell lysate
    Lane 2 : Jurkat (Human T cell leukemia cells from peripheral blood) whole cell lysate
    Lane 3 : HepG2 (Human liver hepatocellular carcinoma) whole cell lysate
    Lane 4 : NCCIT (Human pluripotent embryonic carcinoma) 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: 132 kDa
    Observed band size: 160,170 kDa
    why is the actual band size different from the predicted?


    Exposure time: 30 seconds


    Blocking/Dilution buffer: 5% NFDM/TBST.

    The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID 16702210).

  • Anti-EHMT2/G9A antibody [EPR18894] (ab185050) at 1/1000 dilution + Human fetal heart lysate at 10 µg

    Secondary
    Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/10000 dilution

    Predicted band size: 132 kDa
    Observed band size: 160,170 kDa why is the actual band size different from the predicted?


    Exposure time: 3 minutes


    Blocking/Dilution buffer: 5% NFDM/TBST.

    The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID 16702210).

  • Anti-EHMT2/G9A antibody [EPR18894] (ab185050) at 1/1000 dilution + Human fetal kidney lysate at 10 µg

    Secondary
    Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/10000 dilution

    Predicted band size: 132 kDa
    Observed band size: 160,170 kDa why is the actual band size different from the predicted?


    Exposure time: 8 seconds


    Blocking/Dilution buffer: 5% NFDM/TBST.

    The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID 16702210).

  • All lanes : Anti-EHMT2/G9A antibody [EPR18894] (ab185050) at 1/1000 dilution

    Lane 1 : F9 (Mouse embyro testicular cancer cell line) whole cell lysate
    Lane 2 : Neuro-2a (Mouse neuroblastoma cells) whole cell lysate
    Lane 3 : LLC1 (Mouse lung carcinoma 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

    Predicted band size: 132 kDa
    Observed band size: 160,170 kDa why is the actual band size different from the predicted?


    Exposure time: 5 seconds


    Blocking/Dilution buffer: 5% NFDM/TBST.

    The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID 16702210).

  • All lanes : Anti-EHMT2/G9A antibody [EPR18894] (ab185050) at 1/1000 dilution

    Lane 1 : C6 (Rat glial tumor cells) whole cell lysate
    Lane 2 : RAW 264.7 (Mouse macrophage cells transformed with Abelson murine leukemia virus) whole cell lysate
    Lane 3 : PC-12 (Rat adrenal gland pheochromocytoma) whole cell lysate
    Lane 4 : NIH/3T3 (Mouse embyro fibroblast cells) 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

    Predicted band size: 132 kDa
    Observed band size: 160,170 kDa why is the actual band size different from the predicted?


    Exposure time: 8 seconds


    Blocking/Dilution buffer: 5% NFDM/TBST.

    The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID 16702210).

  • 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.

  • 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.

  • 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.

  • 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.

  • 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.

  • 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.

References

This product has been referenced in:
  • Xiong H  et al. Inhibition of Histone Methyltransferase G9a Attenuates Noise-Induced Cochlear Synaptopathy and Hearing Loss. J Assoc Res Otolaryngol N/A:N/A (2019). Read more (PubMed: 30710318) »
  • Deneault E  et al. CNTN5-/+or EHMT2-/+human iPSC-derived neurons from individuals with autism develop hyperactive neuronal networks. Elife 8:N/A (2019). Read more (PubMed: 30747104) »
See all 3 Publications for this product

Customer reviews and Q&As

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1-4 of 4 Abreviews

Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (Human Dermal Microvascular Endothelial Cells)
Permeabilization
Yes - 0.1% Triton-X100
Specification
Human Dermal Microvascular Endothelial Cells
Blocking step
BSA as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 25°C
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted Jul 14 2017

Application
Immunoprecipitation
Sample
Human Cell lysate - whole cell (Dermal microvascular endothelium)
Total protein in input
200 µg
Immuno-precipitation step
Protein A/G
Specification
Dermal microvascular endothelium

Abcam user community

Verified customer

Submitted Apr 20 2017

Application
ChIP
Sample
Human Cell lysate - nuclear (B Cells)
Negative control
Isotype IgG
Specification
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: Methanol free Formaldehyde
Positive control
Gene promoter specific primers

Abcam user community

Verified customer

Submitted Mar 17 2017

Application
Western blot
Sample
Human Cell lysate - nuclear (Human Dermal Microvascular Endothelial Cells)
Gel Running Conditions
Reduced Denaturing (7.5%)
Loading amount
20 µg
Specification
Human Dermal Microvascular Endothelial Cells
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Feb 28 2017

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