Anti-EED antibody (ab4469)
Key features and details
- Rabbit polyclonal to EED
- Suitable for: WB, ICC/IF
- Knockout validated
- Reacts with: Human, Recombinant fragment
- Isotype: IgG
Overview
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Product name
Anti-EED antibody
See all EED primary antibodies -
Description
Rabbit polyclonal to EED -
Host species
Rabbit -
Specificity
ab4469 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 Applications & Species
See all applications and species dataApplication Species ICC/IF HumanWB HumanRecombinant fragment
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Immunogen
Synthetic peptide corresponding to Human EED aa 1-100 conjugated to keyhole limpet haemocyanin.
(Peptide available asab13743) -
Positive control
- This antibody gave a positive signal in Human Recombinant EED Protein
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.40
Preservative: 0.02% Sodium azide
Constituent: PBS
Batches of this product that have a concentration
Concentration information loading...Purity
Immunogen affinity purifiedClonality
PolyclonalIsotype
IgGResearch areas
Associated products
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Compatible Secondaries
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Isotype control
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Recombinant Protein
Applications
The Abpromise guarantee
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.
GuaranteedTested applications are guaranteed to work and covered by our Abpromise guarantee.
PredictedPredicted to work for this combination of applications and species but not guaranteed.
IncompatibleDoes not work for this combination of applications and species.
Application Species ICC/IF HumanWB HumanRecombinant fragmentAll applications MouseChickenCowXenopus laevisApplication Abreviews Notes WB (3) 1/200. Predicted molecular weight: 49 kDa.ICC/IF 1/100.Notes WB
1/200. Predicted molecular weight: 49 kDa.ICC/IF
1/100.Target
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Function
Polycomb 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 specificity
Expressed 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 similarities
Belongs to the WD repeat ESC family.
Contains 7 WD repeats. -
Developmental stage
Expression peaks at the G1/S phase boundary. -
Cellular localization
Nucleus. Chromosome. Transiently colocalizes with XIST at inactive X chromosomes. - Information by UniProt
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Database links
- Entrez Gene: 426381 Chicken
- Entrez Gene: 404183 Cow
- Entrez Gene: 8726 Human
- Entrez Gene: 13626 Mouse
- Entrez Gene: 398420 Xenopus laevis
- Omim: 605984 Human
- SwissProt: Q5ZKH3 Chicken
- SwissProt: Q3SZ25 Cow
see all -
Alternative names
- eed antibody
- EED protein antibody
- EED, mouse, homolog of antibody
see all
Images
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Western blot - Anti-EED antibody (ab4469)
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 in wild-type HAP1 cells along with additional cross-reactive bands. No band was observed when EED knockout samples were examined. 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) 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. -
Immunocytochemistry/ Immunofluorescence - Anti-EED antibody (ab4469)Rabbit polyclonal to EED (ab4469) on U-2 OS (Human bone osteosarcoma epithelial cell line) cells. Antibody used at 1/100.
U-2 OS 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). -
Western blot - Anti-EED antibody (ab4469)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).
Protocols
Datasheets and documents
References (24)
ab4469 has been referenced in 24 publications.
- Lee JW et al. RUNX3 regulates cell cycle-dependent chromatin dynamics by functioning as a pioneer factor of the restriction-point. Nat Commun 10:1897 (2019). PubMed: 31015486
- Pandya P et al. PICOT binding to chromatin-associated EED negatively regulates cyclin D2 expression by increasing H3K27me3 at the CCND2 gene promoter. Cell Death Dis 10:685 (2019). PubMed: 31527584
- Wang J et al. Asymmetric Expression of LincGET Biases Cell Fate in Two-Cell Mouse Embryos. Cell 175:1887-1901.e18 (2018). PubMed: 30550787
- Wan L et al. Phosphorylation of EZH2 by AMPK Suppresses PRC2 Methyltransferase Activity and Oncogenic Function. Mol Cell 69:279-291.e5 (2018). PubMed: 29351847
- Anwar T et al. p38-mediated phosphorylation at T367 induces EZH2 cytoplasmic localization to promote breast cancer metastasis. Nat Commun 9:2801 (2018). PubMed: 30022044
Images
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Western blot - Anti-EED antibody (ab4469)
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 in wild-type HAP1 cells along with additional cross-reactive bands. No band was observed when EED knockout samples were examined. 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) 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. -
Immunocytochemistry/ Immunofluorescence - Anti-EED antibody (ab4469)
Rabbit polyclonal to EED (ab4469) on U-2 OS (Human bone osteosarcoma epithelial cell line) cells. Antibody used at 1/100.
U-2 OS 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). -
Western blot - Anti-EED antibody (ab4469)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).
