For the best experience on the Abcam website please upgrade to a modern browser such as Google Chrome

Hello. We're improving abcam.com and we'd welcome your feedback.

Hello. We're improving abcam.com and we'd welcome your feedback.

Infomation icon

We haven't added this to the BETA yet

New BETA website

New BETA website

Hello. We're improving abcam.com and we'd welcome your feedback.

Take a look at our BETA site and see what we’ve done so far.

Switch on our new BETA site

Now available

Search and browse selected products

  • A selection of primary antibodies

Purchase these through your usual distributor

In the coming months

  • Additional product types
  • Supporting content
  • Sign in to your account
  • Purchase online
United States
Your country/region is currently set to:

If incorrect, please enter your country/region into the box below, to view site information related to your country/region.

Call (888) 77-ABCAM (22226) or contact us
Need help? Contact us

  • My account
  • Sign out
Sign in or Register with us

Welcome

Sign in or

Don't have an account?

Register with us
My basket
Quick order
Abcam homepage

  • Research Products
    By product type
    Primary antibodies
    Secondary antibodies
    ELISA and Matched Antibody Pair Kits
    Cell and tissue imaging tools
    Cellular and biochemical assays
    Proteins and Peptides
    By product type
    Proteomics tools
    Agonists, activators, antagonists and inhibitors
    Cell lines and Lysates
    Multiplex miRNA assays
    Multiplex Assays
    By research area
    Cancer
    Cardiovascular
    Cell Biology
    Epigenetics
    Metabolism
    Developmental Biology
    By research area
    Immunology
    Microbiology
    Neuroscience
    Signal Transduction
    Stem Cells
  • Customized Products & Partnerships
    Customized Products & Partnerships

    Customized products and commercial partnerships to accelerate your diagnostic and therapeutic programs.

    Customized products

    Partner with us

  • Support
    Support hub

    Access advice and support for any research roadblock

    View support hub

    Protocols

    Your experiments laid out step by step

    View protocols

  • Events
    • Conference calendar
    • Cancer
    • Cardiovascular
    • Epigenetics & Nuclear signaling
    • Immunology
    • Neuroscience
    • Stem cells
    • Tradeshows
    • Scientific webinars
    Keep up to date with the latest events

    Full event breakdown with abstracts, speakers, registration and more

    View global event calendar

  • Pathways
    Cell signalling pathways

    View all pathways

    View all interactive pathways

  1. Link

    products/primary-antibodies/ehmt2g9a-antibody-epr18894-chip-grade-ab185050.pdf

  1. Send me a copy of this email
    I agree to the terms and conditions.
Epigenetics and Nuclear Signaling Chromatin Modifying Enzymes Methylation
Share by email
Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-EHMT2/G9A antibody [EPR18894] - ChIP Grade (ab185050)

  • Datasheet
  • SDS
  • Certificate of Compliance
Reviews (4) Submit a question References (17)

Product price, shipping and contact information

Currently unavailable

Sorry, we can't display this right now.

Please contact us to place your order, or try again later.

 

Loading size & price…

 

Shipping and order information

Shipping info

Promotion Information

Abpromise

Guaranteed product quality, expert customer support.

Find out more.

Western blot - Anti-EHMT2/G9A antibody [EPR18894] - ChIP Grade (ab185050)
  • Immunocytochemistry/ Immunofluorescence - Anti-EHMT2/G9A antibody [EPR18894] - ChIP Grade (ab185050)
  • ChIP-sequencing - Anti-EHMT2/G9A antibody [EPR18894] - ChIP Grade (ab185050)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-EHMT2/G9A antibody [EPR18894] - ChIP Grade (ab185050)
  • Western blot - Anti-EHMT2/G9A antibody [EPR18894] - ChIP Grade (ab185050)
  • Western blot - Anti-EHMT2/G9A antibody [EPR18894] - ChIP Grade (ab185050)
  • Western blot - Anti-EHMT2/G9A antibody [EPR18894] - ChIP Grade (ab185050)
  • Western blot - Anti-EHMT2/G9A antibody [EPR18894] - ChIP Grade (ab185050)
  • Western blot - Anti-EHMT2/G9A antibody [EPR18894] - ChIP Grade (ab185050)
  • Western blot - Anti-EHMT2/G9A antibody [EPR18894] - ChIP Grade (ab185050)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-EHMT2/G9A antibody [EPR18894] - ChIP Grade (ab185050)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-EHMT2/G9A antibody [EPR18894] - ChIP Grade (ab185050)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-EHMT2/G9A antibody [EPR18894] - ChIP Grade (ab185050)
  • Immunocytochemistry/ Immunofluorescence - Anti-EHMT2/G9A antibody [EPR18894] - ChIP Grade (ab185050)
  • Flow Cytometry (Intracellular) - Anti-EHMT2/G9A antibody [EPR18894] - ChIP Grade (ab185050)
  • ChIC/CUT&RUN sequencing - Anti-EHMT2/G9A antibody [EPR18894] - ChIP Grade (ab185050)
  • Immunoprecipitation - Anti-EHMT2/G9A antibody [EPR18894] - ChIP Grade (ab185050)
  • Anti-EHMT2/G9A antibody [EPR18894] - ChIP Grade (ab185050)

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 - ChIP Grade
  • Suitable for: ChIP-sequencing, WB, IHC-P, ICC/IF, IP, Flow Cyt (Intra), ChIC/CUT&RUN-seq
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

Conjugates logo Related conjugates and formulations

Alexa Fluor® 488 Carrier Free

You may also be interested in

Preparation
Product image
Nuclear Extraction Kit (ab113474)
Primary
Product image
Anti-Aly/Ref antibody [EPR17942] (ab202894)
Secondary
Product image
Goat Anti-Rabbit IgG H&L (HRP) (ab205718)

View more associated products

Overview

  • Product name

    Anti-EHMT2/G9A antibody [EPR18894] - ChIP Grade
    See all EHMT2/G9A primary antibodies
  • Description

    Rabbit monoclonal [EPR18894] to EHMT2/G9A - ChIP Grade
  • Host species

    Rabbit
  • Tested applications

    Suitable for: ChIP-sequencing, WB, IHC-P, ICC/IF, IP, Flow Cyt (Intra), ChIC/CUT&RUN-seqmore 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, 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 (intra): HeLa cells. IP: HeLa whole cell lysate. ChIP-seq: HeLa cells. ChIC/CUT&RUN-seq: wild-type HeLa cell line (ab255928).
  • General notes

    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

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
  • Storage buffer

    pH: 7.2
    Preservative: 0.01% Sodium azide
    Constituents: 59% PBS, 40% Glycerol, 0.05% BSA
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR18894
  • Isotype

    IgG
  • Research areas

    • Epigenetics and Nuclear Signaling
    • Chromatin Modifying Enzymes
    • Methylation
    • Stem Cells
    • Embryonic Stem Cells
    • Intracellular
    • Stem Cells
    • Germline Stem Cells
    • Embryonic Germ Cells
    • Epigenetics and Nuclear Signaling
    • Chromatin Modifying Enzymes
    • Methylation
    • Lysine methylation
    • Epigenetics and Nuclear Signaling
    • ChIP assays
    • ChIP antibodies
    • Developmental Biology
    • Reproduction
    • Germ cell markers
    • Developmental Biology
    • Embryogenesis
    • Embryonic stem cells
    • Intracellular

Associated products

  • Alternative Versions

    • Alexa Fluor® 488 Anti-EHMT2/G9A antibody [EPR18894] (ab218359)
    • Anti-EHMT2/G9A antibody [EPR18894] - ChIP Grade - BSA and Azide free (ab240289)
  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
    • Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab97051)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • KO cell lines

    • Human EHMT2 (G9A) knockout HeLa cell line (ab265149)
  • KO cell lysates

    • Human EHMT2 (G9A) knockout HeLa cell lysate (ab257080)
  • Recombinant Protein

    • Recombinant human EHMT2/G9A protein (ab80323)
  • Related Products

    • VeriBlot for IP Detection Reagent (HRP) (ab131366)

Applications

The Abpromise guarantee

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
ChIP-sequencing
Use 8µg for 107 cells.
WB (1)
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)
1/1000.
IP (1)
1/50.
Flow Cyt (Intra)
1/150.
ChIC/CUT&RUN-seq
Use at an assay dependent concentration.
Notes
ChIP-sequencing
Use 8µg for 107 cells.
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.
IP
1/50.
Flow Cyt (Intra)
1/150.
ChIC/CUT&RUN-seq
Use at an assay dependent concentration.

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.
  • Target information above from: UniProt accession Q96KQ7 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 10919 Human
    • Entrez Gene: 110147 Mouse
    • Entrez Gene: 361798 Rat
    • Omim: 604599 Human
    • SwissProt: Q96KQ7 Human
    • SwissProt: Q9Z148 Mouse
    • Unigene: 709218 Human
    • Unigene: 35345 Mouse
    • 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] - ChIP Grade (ab185050)
      Western blot - Anti-EHMT2/G9A antibody [EPR18894] - ChIP Grade (ab185050)
      All lanes : Anti-EHMT2/G9A antibody [EPR18894] - ChIP Grade (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



      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.

       

    • Immunocytochemistry/ Immunofluorescence - Anti-EHMT2/G9A antibody [EPR18894] - ChIP Grade (ab185050)
      Immunocytochemistry/ Immunofluorescence - Anti-EHMT2/G9A antibody [EPR18894] - ChIP Grade (ab185050)

      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.

    • ChIP-sequencing - Anti-EHMT2/G9A antibody [EPR18894] - ChIP Grade (ab185050)
      ChIP-sequencing - Anti-EHMT2/G9A antibody [EPR18894] - ChIP Grade (ab185050)

      Chromatin was prepared from HeLa cells. Cells were fixed with 1% formaldehyde for 10 minutes. ChIP was performed with 107 HeLa cells and 8 µg of ab185050 [EPR18894]. ChIP DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 30 million reads.

      Additional screenshots of mapped reads can be downloaded here.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-EHMT2/G9A antibody [EPR18894] - ChIP Grade (ab185050)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-EHMT2/G9A antibody [EPR18894] - ChIP Grade (ab185050)

      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.

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

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

      Lane 1 : Wild-type HAP1 cell lysate
      Lane 2 : EHMT2/G9A knockout HAP1 cell lysate
      Lane 3 : HEK293 cell lysate
      Lane 4 : HepG2 cell lysate

      Lysates/proteins at 20 µg per lane.

      Predicted band size: 132 kDa



      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.

    • Western blot - Anti-EHMT2/G9A antibody [EPR18894] - ChIP Grade (ab185050)
      Western blot - Anti-EHMT2/G9A antibody [EPR18894] - ChIP Grade (ab185050)
      All lanes : Anti-EHMT2/G9A antibody [EPR18894] - ChIP Grade (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).

    • Western blot - Anti-EHMT2/G9A antibody [EPR18894] - ChIP Grade (ab185050)
      Western blot - Anti-EHMT2/G9A antibody [EPR18894] - ChIP Grade (ab185050)
      Anti-EHMT2/G9A antibody [EPR18894] - ChIP Grade (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).

    • Western blot - Anti-EHMT2/G9A antibody [EPR18894] - ChIP Grade (ab185050)
      Western blot - Anti-EHMT2/G9A antibody [EPR18894] - ChIP Grade (ab185050)
      Anti-EHMT2/G9A antibody [EPR18894] - ChIP Grade (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).

    • Western blot - Anti-EHMT2/G9A antibody [EPR18894] - ChIP Grade (ab185050)
      Western blot - Anti-EHMT2/G9A antibody [EPR18894] - ChIP Grade (ab185050)
      All lanes : Anti-EHMT2/G9A antibody [EPR18894] - ChIP Grade (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).

    • Western blot - Anti-EHMT2/G9A antibody [EPR18894] - ChIP Grade (ab185050)
      Western blot - Anti-EHMT2/G9A antibody [EPR18894] - ChIP Grade (ab185050)
      All lanes : Anti-EHMT2/G9A antibody [EPR18894] - ChIP Grade (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).

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-EHMT2/G9A antibody [EPR18894] - ChIP Grade (ab185050)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-EHMT2/G9A antibody [EPR18894] - ChIP Grade (ab185050)

      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.

      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] - ChIP Grade (ab185050)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-EHMT2/G9A antibody [EPR18894] - ChIP Grade (ab185050)

      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.

      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] - ChIP Grade (ab185050)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-EHMT2/G9A antibody [EPR18894] - ChIP Grade (ab185050)

      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.

      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] - ChIP Grade (ab185050)
      Immunocytochemistry/ Immunofluorescence - Anti-EHMT2/G9A antibody [EPR18894] - ChIP Grade (ab185050)

      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 Cytometry (Intracellular) - Anti-EHMT2/G9A antibody [EPR18894] - ChIP Grade (ab185050)
      Flow Cytometry (Intracellular) - Anti-EHMT2/G9A antibody [EPR18894] - ChIP Grade (ab185050)

      Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed HeLa (Human epithelial cells from cervix adenocarcinoma) cells labeling EHMT2/G9Awith 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. 

       

       

    • ChIC/CUT&RUN sequencing - Anti-EHMT2/G9A antibody [EPR18894] - ChIP Grade (ab185050)
      ChIC/CUT&RUN sequencing - Anti-EHMT2/G9A antibody [EPR18894] - ChIP Grade (ab185050)

      ChIC/CUT&RUN was performed using the ChIC/CUT&RUN pAG-MNAse ab285373. 2.5X10^5 of wild-type HeLa cell line (ab255928) or EHMT2 (G9A) knockout HeLa cell line (ab265149) were used along with 5µg of Anti-EHMT2/G9A antibody [EPR18894] - ChIP Grade (ab185050). Assay Quality Control was conducted using 5µg Anti-CTCF antibody [EPR18253] - ChIP Grade (ab188408) on the same cell lines. The resulting DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 10 million reads. The Negative IgG control is also shown.

      Additional screenshots of mapped reads can be downloaded here.

    • Immunoprecipitation - Anti-EHMT2/G9A antibody [EPR18894] - ChIP Grade (ab185050)
      Immunoprecipitation - Anti-EHMT2/G9A antibody [EPR18894] - ChIP Grade (ab185050)

      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.

    • Anti-EHMT2/G9A antibody [EPR18894] - ChIP Grade (ab185050)
      Anti-EHMT2/G9A antibody [EPR18894] - ChIP Grade (ab185050)

    Protocols

    • Flow cytometry protocols
    • Immunoprecipitation protocols
    • Immunohistochemistry protocols
    • Immunocytochemistry & immunofluorescence protocols
    • Western blot protocols

    Click here to view the general protocols

    Datasheets and documents

    • SDS download

    • Datasheet download

      Download

    Certificate of Compliance

    To download a Certificate of Compliance, please enter your Lot number below:

    References (17)

    Publishing research using ab185050? Please let us know so that we can cite the reference in this datasheet.

    ab185050 has been referenced in 17 publications.

    • Cipriano A  et al. Epigenetic regulation of Wnt7b expression by the cis-acting long noncoding RNA Lnc-Rewind in muscle stem cells. Elife 10:N/A (2021). PubMed: 33432928
    • Hou QQ  et al. TBC1D3 promotes neural progenitor proliferation by suppressing the histone methyltransferase G9a. Sci Adv 7:N/A (2021). PubMed: 33523893
    • Barcena-Varela M  et al. Epigenetic mechanisms and metabolic reprogramming in fibrogenesis: dual targeting of G9a and DNMT1 for the inhibition of liver fibrosis. Gut 70:388-400 (2021). PubMed: 32327527
    • Wang H  et al. Post-Treatment Sevoflurane Protects Against Hypoxic-Ischemic Brain Injury in Neonatal Rats by Downregulating Histone Methyltransferase G9a and Upregulating Nuclear Factor Erythroid 2-Related Factor 2 (NRF2). Med Sci Monit 27:e930042 (2021). PubMed: 34059615
    • Xia S  et al. SLC7A2 deficiency promotes hepatocellular carcinoma progression by enhancing recruitment of myeloid-derived suppressors cells. Cell Death Dis 12:570 (2021). PubMed: 34108444
    View all Publications for this product

    Customer reviews and Q&As

    Show All Reviews Q&A
    Submit a review Submit a question

    1-4 of 4 Abreviews or Q&A

    Immunocytochemistry/ Immunofluorescence abreview for Anti-EHMT2/G9A antibody [EPR18894] - ChIP Grade

    Excellent
    Abreviews
    Abreviews
    abreview image
    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
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    Abcam user community

    Verified customer

    Submitted Jul 14 2017

    Immunoprecipitation abreview for Anti-EHMT2/G9A antibody [EPR18894] - ChIP Grade

    Good
    Abreviews
    Abreviews
    abreview image
    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
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    Abcam user community

    Verified customer

    Submitted Apr 20 2017

    ChIP abreview for Anti-EHMT2/G9A antibody [EPR18894] - ChIP Grade

    Good
    Abreviews
    Abreviews
    abreview image
    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
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    Abcam user community

    Verified customer

    Submitted Mar 17 2017

    Western blot abreview for Anti-EHMT2/G9A antibody [EPR18894] - ChIP Grade

    Excellent
    Abreviews
    Abreviews
    abreview image
    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
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    Abcam user community

    Verified customer

    Submitted Feb 28 2017

    Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
    For licensing inquiries, please contact partnerships@abcam.com

    Get resources and offers direct to your inbox Sign up
    A-Z by research area
    • Cancer
    • Cardiovascular
    • Cell biology
    • Developmental biology
    • Epigenetics & Nuclear signaling
    • Immunology
    • Metabolism
    • Microbiology
    • Neuroscience
    • Signal transduction
    • Stem cells
    A-Z by product type
    • Primary antibodies
    • Secondary antibodies
    • Biochemicals
    • Isotype controls
    • Flow cytometry multi-color selector
    • Kits
    • Loading controls
    • Lysates
    • Peptides
    • Proteins
    • Slides
    • Tags and cell markers
    • Tools & Reagents
    Help & support
    • Support
    • Make an Inquiry
    • Protocols & troubleshooting
    • Placing an order
    • RabMAb products
    • Biochemical product FAQs
    • Training
    • Browse by Target
    Company
    • Corporate site
    • Investor relations
    • Company news
    • Careers
    • About us
    • Blog
    Events
    • Tradeshows
    • Conferences
    International websites
    • abcam.cn
    • abcam.co.jp

    Join with us

    • LinkedIn
    • facebook
    • Twitter
    • YouTube
    • Terms of sale
    • Website terms of use
    • Cookie policy
    • Privacy policy
    • Legal
    • Modern slavery statement
    © 1998-2023 Abcam plc. All rights reserved.