- Clone
- W18151A (See other available formats)
- Regulatory Status
- RUO
- Other Names
- Embryonic Ectoderm Development, Embryonic Ectoderm Development Protein, Polycomb Protein EED
- Isotype
- Rat IgG2a, κ
- Ave. Rating
- Submit a Review
- Product Citations
- publications
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Whole cell extracts (15 µg total protein) from the indicated cell lines were resolved by 4-12% Bis-Tris gel electrophoresis, transferred to a PVDF membrane, and probed with 1.0 µg/mL of purified anti-EED antibody (clone W18151A) overnight at 4°C. Proteins were visualized by chemiluminescence detection using HRP goat anti-rat IgG antibody (Cat. No. 405405) at a 1:3000 dilution. Direct-Blot™ HRP anti-GAPDH antibody (Cat. No. 607904) was used as a loading control at a 1:50000 dilution (lower). Lane M: Molecular weight marker. -
Whole cell extracts (250 µg total protein) prepared from HeLa cells were immunoprecipitated overnight with 2.5 µg of purified rat IgG2a, κ isotype ctrl antibody (Cat. No. 400502) or purified anti-EED antibody (clone W18151A). The resulting IP fractions and whole cell extract input (5%) were resolved by 4-12% Bis-Tris gel electrophoresis, transferred to a PVDF membrane and probed with 1.0 µg/mL of purified anti-EED antibody (clone W18151A) overnight at 4°C. Proteins were visualized by chemiluminescence detection using HRP anti-rat light chain specific antibody. Direct-Blot™ HRP anti-GAPDH antibody (Cat. No. 607904) was used as a loading control at a 1:50000 dilution (lower). Lane M: Molecular weight marker. -
Whole cell extracts (15 µg protein) from HeLa cells transfected with scramble control siRNA or siRNA targeting EED were resolved on a 4-12% Bis-Tris gel electrophoresis, transferred to a PVDF membrane and probed with 1.0 µg/mL of purified anti-EED antibody (clone W18151A) overnight at 4°C. Proteins were visualized by chemiluminescence detection using HRP goat anti-rat IgG antibody (Cat. No. 405405) at a 1:3000 dilution. Direct-Blot™ HRP anti-GAPDH antibody (Cat. No. 607904) was used as a loading control at a 1:50000 dilution (lower). Lane M: Molecular weight marker. siRNA targeting EED for depletion was obtained from Dharmcon (SMARTpool ON-TARGETplus EED siRNA, Cat. No. L-017581-00-0005) -
HeLa cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with Triton X-100 for 10 minutes, and blocked with 5% FBS for 60 minutes. Cells were then intracellularly stained with a 1 µg/mL of either purified rat IgG2a, κ isotype ctrl antibody (Cat. No. 400502, panel A) or purified anti-EED antibody (clone W18151A, panel B) overnight at 4°C, followed by incubation with Alexa Fluor® 594 goat anti-rat IgG antibody (Cat. No. 405422) at 2.0 µg/mL. Nuclei were counterstained with DAPI and the image was captured with a 60X objective.
Cat # | Size | Price | Quantity Check Availability | Save | ||
---|---|---|---|---|---|---|
934901 | 25 µg | 81€ | ||||
934902 | 100 µg | 193€ |
Embryonic ectoderm development (EED) is a member of the polycomb-group (PcG) family. The PcG family members form multimeric protein complexes called polycomb repressive complexes (PRCs). These polycomb repressive complexes (PRCs) epigenetically silence gene expression and are involved in a variety of biological processes such as cell cycle control, stem cell plasticity, cellular senescence and cancer.
There are multiple isoforms (~4), which seem to interact differently with different PRCs. Isoform 1 of EED also has an alternate phosphorylated form. Diseases associated with EED include Cohen-Gibson syndrome and Weaver syndrome.
Product Details
- Verified Reactivity
- Human, Mouse
- Antibody Type
- Monoclonal
- Host Species
- Rat
- Immunogen
- Partial recombinant human EED protein of approximately 250 amino acids
- Formulation
- Phosphate-buffered solution, pH 7.2, containing 0.09% sodium azide
- Preparation
- The antibody was purified by affinity chromatography.
- Concentration
- 0.5 mg/mL
- Storage & Handling
- The antibody solution should be stored undiluted between 2°C and 8°C.
- Application
-
WB - Quality tested
IP, ICC, KO/KD-WB - Verified - Recommended Usage
-
Each lot of this antibody is quality control tested by western blotting. For western blotting, the suggested use of this reagent is 0.125 - 1.0 µg/mL. For immunoprecipitation, the suggested use of this reagent is 2.5 µg/test. For immunocytochemistry, a concentration range of 1.0 - 5.0 μg/mL is recommended. It is recommended that the reagent be titrated for optimal performance for each application.
- Application Notes
-
Clone recognizes 3 isoforms of human and mouse EED.
For ICC testing, HeLa cells were fixed in 4% PFA and permeabilized with either Triton X-100 or methanol; both methods produced strong staining at 1 mg/mL. - RRID
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AB_2820221 (BioLegend Cat. No. 934901)
AB_2820221 (BioLegend Cat. No. 934902)
Antigen Details
- Structure
- EED is an ~440 amino acid protein with 4 isoforms of a predicted molecular weight of around 50 kD.
- Distribution
-
EED mainly localizes to the nucleoplasm.
- Function
- Maintains transcriptionally repressive state of genes.
- Interaction
- EZH2
- Biology Area
- Cancer Biomarkers, Cell Cycle/DNA Replication, Cell Death, Chromatin Remodeling/Epigenetics, Stem Cells
- Antigen References
-
Van der Vlag J, et al. 1999. Nat. Gen. 23:474.
Martin C, et al. 2006. J. Bol. 281:13:8365.
Cao Q, et al. 2014. Nat. Comm. 5:3127.
Kuzmichev A, et al. 2004. Mol. Cell. 14:183.
Vire E, et al. 2005. Nat. 439:871.
Schuettengruber B, et al. 2009. Development. 136: 353.
- Gene ID
- 8726 View all products for this Gene ID
- UniProt
- View information about EED on UniProt.org
Other Formats
View All EED Reagents Request Custom ConjugationDescription | Clone | Applications |
---|---|---|
Purified anti-EED | W18151A | WB,IP,ICC |
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Your actual data may vary due to variations in samples, target cells, instruments and their settings, staining conditions, and other factors.
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