Anti-EED antibody (ab4469)
- Datasheet
- References (20)
- Protocols
Overview
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Product nameAnti-EED antibody
See all EED primary antibodies -
DescriptionRabbit polyclonal to EED
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Host speciesRabbit
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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.
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Tested applicationsSuitable for: WB, ICC/IFmore details
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Species reactivityReacts with: Cow, Human
Predicted to work with: Mouse, Chicken, Xenopus laevis
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Immunogen
Synthetic peptide conjugated to KLH derived from within residues 1 - 100 of Human EED.
Read Abcam's proprietary immunogen policy (Peptide available as ab13743.) -
Positive control
- This antibody gave a positive signal in Human Recombinant EED Protein
Properties
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FormLiquid
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Storage instructionsShipped 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.
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Storage bufferPreservative: 0.02% Sodium Azide
Constituents: 1% BSA, PBS, pH 7.4 -
Concentration information loading... -
PurityImmunogen affinity purified
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ClonalityPolyclonal
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IsotypeIgG
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Research areas
Associated products
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Compatible Secondaries
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Isotype control
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. | |
| ICC/IF | 1/100. |
Target
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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.
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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.
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Sequence similaritiesBelongs to the WD repeat ESC family.
Contains 7 WD repeats. -
Developmental stageExpression peaks at the G1/S phase boundary.
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Cellular localizationNucleus. 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 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). -
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).
Datasheets and documents
References
This product has been referenced in:
- Anwar T et al. p38-mediated phosphorylation at T367 induces EZH2 cytoplasmic localization to promote breast cancer metastasis. Nat Commun 9:2801 (2018). Read more (PubMed: 30022044) »
- Labade AS et al. HOXA repression is mediated by nucleoporin Nup93 assisted by its interactors Nup188 and Nup205. Epigenetics Chromatin 9:54 (2016). WB ; Human . Read more (PubMed: 27980680) »
