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  1. Link

    kmt6--ezh2-antibody-epr20108-chip-grade-ab191250.pdf

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Epigenetics and Nuclear Signaling Chromatin Modifying Enzymes Methylation
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Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-KMT6 / EZH2 antibody [EPR20108] - ChIP Grade (ab191250)

  • Datasheet
  • SDS
  • Certificate of Compliance
Reviews (5) Submit a question References (27)

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ChIP - Anti-KMT6 / EZH2 antibody [EPR20108] - ChIP Grade (ab191250)
  • Western blot - Anti-KMT6 / EZH2 antibody [EPR20108] - ChIP Grade (ab191250)
  • Western blot - Anti-KMT6 / EZH2 antibody [EPR20108] - ChIP Grade (ab191250)
  • Western blot - Anti-KMT6 / EZH2 antibody [EPR20108] - ChIP Grade (ab191250)
  • Immunoprecipitation - Anti-KMT6 / EZH2 antibody [EPR20108] - ChIP Grade (ab191250)
  • Anti-KMT6 / EZH2 antibody [EPR20108] - ChIP Grade (ab191250)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR20108] to KMT6 / EZH2 - ChIP Grade
  • Suitable for: ChIP, WB, IP
  • Knockout validated
  • Reacts with: Human

Conjugates logo Related conjugates and formulations

Carrier Free

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Overview

  • Product name

    Anti-KMT6 / EZH2 antibody [EPR20108] - ChIP Grade
    See all KMT6 / EZH2 primary antibodies
  • Description

    Rabbit monoclonal [EPR20108] to KMT6 / EZH2 - ChIP Grade
  • Host species

    Rabbit
  • Tested applications

    Suitable for: ChIP, WB, IPmore details
  • Species reactivity

    Reacts with: Human
  • Immunogen

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • WB: MCF7, HeLa, U-2 OS and HEK-293 whole cell lysates; Wild-type HAP1 cell lysate. IP: HeLa whole cell lysate. ChIP: Chromatin from MDA-MB-231 cells.
  • 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

    EPR20108
  • Isotype

    IgG
  • Research areas

    • Epigenetics and Nuclear Signaling
    • Chromatin Modifying Enzymes
    • Methylation
    • Epigenetics and Nuclear Signaling
    • Chromatin Remodeling
    • Polycomb Silencing
    • PRC2
    • Epigenetics and Nuclear Signaling
    • Chromatin Remodeling
    • Polycomb Silencing
    • PRC3
    • Epigenetics and Nuclear Signaling
    • Chromatin Modifying Enzymes
    • Methylation
    • Lysine methylation
    • Cancer
    • Tumor biomarkers
    • Enzymes
    • Other
    • Epigenetics and Nuclear Signaling
    • ChIP assays
    • ChIP antibodies

Associated products

  • Alternative Versions

    • Anti-KMT6 / EZH2 antibody [EPR20108] - BSA and Azide free (ab240992)
  • 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 (HRP) (ab97051)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • Recombinant Protein

    • Recombinant Human KMT6 / EZH2 protein (ab132934)
  • Related Products

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

Applications

The Abpromise guarantee

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
ChIP (2)
Use 5 µg for 25 µg of chromatin.
WB (2)
1/1000. Detects a band of approximately 90 kDa (predicted molecular weight: 85 kDa).
IP
1/30.
Notes
ChIP
Use 5 µg for 25 µg of chromatin.
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.
  • Target information above from: UniProt accession Q15910 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 2146 Human
    • Omim: 601573 Human
    • SwissProt: Q15910 Human
    • Unigene: 444082 Human
    • 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

    • ChIP - Anti-KMT6 / EZH2 antibody [EPR20108] - ChIP Grade (ab191250)
      ChIP - Anti-KMT6 / EZH2 antibody [EPR20108] - ChIP Grade (ab191250)

      Chromatin was prepared from MDA-MB-231 cells according to the Abcam X-ChIP protocol. Cells were fixed with formaldehyde for 10 minutes. The ChIP was performed with 25µg of chromatin, 5µg of ab191250 (red), and 20µl of Anti rabbit IgG sepharose beads. 5μg of rabbit normal IgG was added to the beads control (grey).  The immunoprecipitated DNA was quantified by real time PCR (Sybr green approach).

      Cell line and primers selection based on literature (PMID: 21880597).

    • Western blot - Anti-KMT6 / EZH2 antibody [EPR20108] - ChIP Grade (ab191250)
      Western blot - Anti-KMT6 / EZH2 antibody [EPR20108] - ChIP Grade (ab191250)
      All lanes : Anti-KMT6 / EZH2 antibody [EPR20108] - ChIP Grade (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 : MCF7 (human breast adenocarcinoma 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.

    • Western blot - Anti-KMT6 / EZH2 antibody [EPR20108] - ChIP Grade (ab191250)
      Western blot - Anti-KMT6 / EZH2 antibody [EPR20108] - ChIP Grade (ab191250)
      All lanes : Anti-KMT6 / EZH2 antibody [EPR20108] - ChIP Grade (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.

    • Western blot - Anti-KMT6 / EZH2 antibody [EPR20108] - ChIP Grade (ab191250)
      Western blot - Anti-KMT6 / EZH2 antibody [EPR20108] - ChIP Grade (ab191250)
      Anti-KMT6 / EZH2 antibody [EPR20108] - ChIP Grade (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.

    • Immunoprecipitation - Anti-KMT6 / EZH2 antibody [EPR20108] - ChIP Grade (ab191250)
      Immunoprecipitation - Anti-KMT6 / EZH2 antibody [EPR20108] - ChIP Grade (ab191250)

      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.

    • Anti-KMT6 / EZH2 antibody [EPR20108] - ChIP Grade (ab191250)
      Anti-KMT6 / EZH2 antibody [EPR20108] - ChIP Grade (ab191250)

    Protocols

    • ChIP protocols
    • Western blot protocols
    • Immunoprecipitation 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 (27)

    Publishing research using ab191250? Please let us know so that we can cite the reference in this datasheet.

    ab191250 has been referenced in 27 publications.

    • Li R  et al. LINC01116 Promotes Doxorubicin Resistance in Osteosarcoma by Epigenetically Silencing miR-424-5p and Inducing Epithelial-Mesenchymal Transition. Front Pharmacol 12:632206 (2021). PubMed: 33762953
    • Kuang Y  et al. Inhibition of microRNA let-7b expression by KDM2B promotes cancer progression by targeting EZH2 in ovarian cancer. Cancer Sci 112:231-242 (2021). PubMed: 33091189
    • Qu S  et al. EZH2 is involved in psoriasis progression by impairing miR-125a-5p inhibition of SFMBT1 and leading to inhibition of the TGFß/SMAD pathway. Ther Adv Chronic Dis 12:2040622320987348 (2021). PubMed: 33948156
    • Yang L  et al. PRMT5 functionally associates with EZH2 to promote colorectal cancer progression through epigenetically repressing CDKN2B expression. Theranostics 11:3742-3759 (2021). PubMed: 33664859
    • Liu Z  et al. EZH2-mediated epigenetic suppression of SH3BGRL potently inhibits lung cancer progression. Biochem Biophys Res Commun 548:53-59 (2021). PubMed: 33631674
    View all Publications for this product

    Customer reviews and Q&As

    Show All Reviews Q&A
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    1-5 of 5 Abreviews

    ChIP-sequencing abreview for Anti-KMT6 / EZH2 antibody [EPR20108] - ChIP Grade

    Below Average
    Abreviews
    Abreviews
    Application
    ChIP-sequencing
    Sample
    Human Cell lysate - nuclear (K562)
    Specification
    K562
    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.

    MR. Andrew Perez

    Verified customer

    Submitted Mar 24 2022

    ChIP abreview for Anti-KMT6 / EZH2 antibody [EPR20108] - ChIP Grade

    Inconclusive
    Abreviews
    Abreviews
    abreview image
    Application
    ChIP
    Sample
    Mouse Cell lysate - nuclear (Cell line NIH 3T3)
    Negative control
    Aprt, Atp5
    Specification
    Cell line NIH 3T3
    Detection step
    Real-time PCR
    Type
    Cross-linking (X-ChIP)
    Duration of cross-linking step: 10 minute(s) and 0 second(s)
    Specification of the cross-linking agent: 1% PFA
    Positive control
    Sfrp4, Oct4
    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.

    DR. Amalia Tabib

    Verified customer

    Submitted Jan 28 2022

    Western blot abreview for Anti-KMT6 / EZH2 antibody [EPR20108] - ChIP Grade

    Good
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (Smooth Muscle Cells)
    Gel Running Conditions
    Non-reduced Denaturing (1-15%)
    Loading amount
    15 µg
    Specification
    Smooth Muscle Cells
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: rt°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 Nov 26 2021

    Western blot abreview for Anti-KMT6 / EZH2 antibody [EPR20108] - ChIP Grade

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (HEK293T)
    Gel Running Conditions
    Reduced Denaturing (Bolt 4-12% Bis-Tris)
    Loading amount
    33 µg
    Treatment
    DMSO/UNC6852 for 48hrs
    Specification
    HEK293T
    Blocking step
    Intercept (PBS) Blocking Buffer as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 50% · Temperature: ~22°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 Apr 14 2021

    ChIP abreview for Anti-KMT6 / EZH2 antibody [EPR20108]

    Excellent
    Abreviews
    Abreviews
    abreview image
    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
    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 05 2017

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