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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

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 & Species

    Application Species
    ChIP
    Human
    IHC-Fr
    Human
    IP
    Human
    WB
    Human
    See all applications and species data

  • 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

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).

    ChIP was performed according to the 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)

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
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