Overview

  • Product nameAnti-EED antibody
    See all EED primary antibodies
  • Description
    Rabbit polyclonal to EED
  • Specificityab4469 specifically recognises the four isoforms of EED following the immunoprecipitation of tagged Ezh2 from HEK293 nuclear extract. However, using HEK293 nuclear lysate ab4469 predominantly recognises non-specific bands.
  • Tested applicationsSuitable for: WB, IP, ICC/IF, IHC-FoFrmore details
  • Species reactivity
    Reacts with: Cow, Human
    Predicted to work with: Mouse, Chicken, Xenopus laevis
  • Immunogen

    Synthetic peptide conjugated to KLH derived from within residues 1 - 100 of Human EED.

    (Peptide available as ab13743.)

  • Positive control
    • This antibody gave a positive signal in Human Recombinant EED Protein

Properties

Applications

Our Abpromise guarantee covers the use of ab4469 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 1/200. Predicted molecular weight: 49 kDa.
IP Use at an assay dependent concentration.
ICC/IF 1/100.
IHC-FoFr Use at an assay dependent concentration.

Target

  • FunctionPolycomb group (PcG) protein. Component 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. 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 and CDKN2A.
  • Tissue specificityExpressed in brain, colon, heart, kidney, liver, lung, muscle, ovary, peripheral blood leukocytes, pancreas, placenta, prostate, spleen, small intestine, testis, thymus and uterus. Appears to be overexpressed in breast and colon cancer.
  • Sequence similaritiesBelongs to the WD repeat ESC family.
    Contains 7 WD repeats.
  • Developmental stageExpression peaks at the G1/S phase boundary.
  • Cellular localizationNucleus. Chromosome. Transiently colocalizes with XIST at inactive X chromosomes.
  • Information by UniProt
  • Database links
  • Alternative names
    • eed antibody
    • EED protein antibody
    • EED, mouse, homolog of antibody
    • EED_HUMAN antibody
    • Embryonic ectoderm development antibody
    • Embryonic ectoderm development isoform a antibody
    • Embryonic ECTODERM development protein homolog antibody
    • Embryonic ectoderm development protein, mouse, homolog of antibody
    • hEED antibody
    • OTTHUMP00000235483 antibody
    • OTTHUMP00000235484 antibody
    • OTTHUMP00000235485 antibody
    • Polycomb protein eed antibody
    • WAIT 1 antibody
    • WAIT-1 antibody
    • WAIT1 antibody
    • WD protein associating with integrin cytoplasmic tails 1 antibody
    see all

Anti-EED antibody images



  • Predicted band size : 49 kDa

    Lane 1: Wild-type HAP1 cell lysate (20 µg)
    Lane 2: EED knockout HAP1 cell lysate (20 µg)
    Lane 3: HeLa cell lysate (20 µg)
    Lane 4: HepG2 cell lysate (20 µg)

    Lanes 1 - 4: Merged signal (red and green). Green - ab4469 observed at 55 kDa. Red - loading control, ab8245, observed at 37 kDa.

    ab4469 was shown to recognize EED when EED knockout samples were used, along with additional cross-reactive bands. Wild-type and EED knockout samples were subjected to SDS-PAGE. ab4469 and ab8245 (loading control to GAPDH) were diluted 1/250 and 1/1000 respectively and incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) ab216776 secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.

  • Rabbit polyclonal to EED (ab4469) on U20S cells. Antibody used at 1/100.

    U20S cells were fixed with PFA (3.7%), permeabilised with PBS 0.1% Triton. Counterstained with Hoechst. Cells were examined with Axioplan 2 microscope (Zeiss).

    Secondary antibody: donkey anti-rabbit (Cy3).



  • Predicted band size : 49 kDa

    Rabbit polyclonal to EED (ab4469) at 1/200.

    Lane 1: WB on HeLa nuclear extract (250ug).

    Lanes 2-5: IP - U20S cells were transfected with 4ug of HA-EED. WB using TBS milk 2%, BSA 0.5%.

     

    Rabbit polyclonal to EED (ab4469) at 1/200.
    Lane 1: WB on HeLa nuclear extract (250ug).

    Lanes 2-5: IP - U20S cells were transfected with 4ug of HA-EED. WB using TBS milk 2%, BSA 0.5%.

     

    Secondary antibody anti-rabbit HRP

  • Anti-EED antibody (ab4469) at 1/250 dilution + EED Recombinant Protein (Human) at 0.1 µg

    Secondary
    Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution

    Performed under reducing conditions.

    Predicted band size : 49 kDa
    Observed band size : 80 kDa (why is the actual band size different from the predicted?)
    ab4469 recognizes the full-length recombinant EED protein which has an expected molecular weight of 80 kDa (tagged).

References for Anti-EED antibody (ab4469)

This product has been referenced in:
  • Mitic T  et al. EZH2 modulates angiogenesis in vitro and in a mouse model of limb ischemia. Mol Ther 23:32-42 (2015). Read more (PubMed: 25189741) »
  • Gonzalez ME  et al. EZH2 expands breast stem cells through activation of NOTCH1 signaling. Proc Natl Acad Sci U S A 111:3098-103 (2014). WB ; Human . Read more (PubMed: 24516139) »

See all 13 Publications for this product

Product Wall

Abreviews
Application Western blot
Loading amount 30 µg
Gel Running Conditions Non-reduced Denaturing (10%)
Sample Mouse Cell lysate - whole cell (Extra Embryonic Endoderm Cells (XEN))
Specification Extra Embryonic Endoderm Cells (XEN)
Blocking step BSA as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 25°C
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Submitted Nov 27 2013

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