Rabbit Recombinant Monoclonal EED antibody. Suitable for IP, ChIP, WB, ChIP-seq and reacts with Human, Mouse samples. Cited in 2 publications.
IgG
Rabbit
Preservative: 0.01% Sodium azide
Constituents: 59.94% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Liquid
Monoclonal
IP | ChIP | WB | IHC-P | ICC/IF | ChIP-seq | Flow Cyt | |
---|---|---|---|---|---|---|---|
Human | Tested | Tested | Tested | Not recommended | Not recommended | Expected | Not recommended |
Mouse | Expected | Expected | Tested | Not recommended | Not recommended | Tested | Not recommended |
Rat | Not recommended | Not recommended | Not recommended | Not recommended | Not recommended | Not recommended | Not recommended |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info 1/30 | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Mouse | Dilution info Use at an assay dependent concentration. | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Rat | Dilution info - | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info 5 µg chromatin for 25.00000 µg chromatin | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Mouse | Dilution info Use at an assay dependent concentration. | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Rat | Dilution info - | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Mouse | Dilution info 1/1000 | Notes - |
Species Human | Dilution info 1/1000 | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Rat | Dilution info - | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Mouse, Human, Rat | Dilution info - | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Mouse, Human, Rat | Dilution info - | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Mouse | Dilution info 0.1 µL for 1.00000 µg chromatin | Notes Use at 0.1 uL/ug chromatin. |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info Use at an assay dependent concentration. | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Rat | Dilution info - | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Human, Mouse, Rat | Dilution info - | Notes - |
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. Also recognizes 'Lys-26' trimethylated histone H1 with the effect of inhibiting PRC2 complex methyltransferase activity on nucleosomal histone H3 'Lys-27', whereas H3 'Lys-27' recognition has the opposite effect, enabling the propagation of this repressive mark. 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.
Polycomb protein EED, hEED, Embryonic ectoderm development protein, WD protein associating with integrin cytoplasmic tails 1, WAIT-1, EED
Rabbit Recombinant Monoclonal EED antibody. Suitable for IP, ChIP, WB, ChIP-seq and reacts with Human, Mouse samples. Cited in 2 publications.
IgG
Rabbit
Preservative: 0.01% Sodium azide
Constituents: 59.94% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Liquid
Monoclonal
EPR23043-5
Affinity purification Protein A
ab240650 detects an unknown band close to the target bands in cytoplasm.
Blue Ice
1-2 weeks
+4°C
-20°C
Upon delivery aliquot
Avoid freeze / thaw cycle
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
This product is a recombinant monoclonal antibody, which offers several advantages including:
For more information, read more on recombinant antibodies.
This supplementary information is collated from multiple sources and compiled automatically.
The EED protein also known as embryonic ectoderm development plays an important mechanical role within the cell. It is a component of the polycomb repressive complex 2 (PRC2) that mediates transcriptional silencing of target genes. The molecular weight of EED protein is approximately 50 kDa. EED is expressed in various tissues and is a critical mediator in maintaining transcriptional repression necessary for embryonic stem cell identity and differentiation.
EED functions as a repressor of gene transcription through the polycomb group complex. It interacts with other components of PRC2 such as EZH2 SUZ12 to establish and maintain chromatin modifications. EED acts as a scaffold allowing communication between other proteins essential in the assembly and function of PRC2. These interactions regulate histone methylation contributing to gene silencing.
EED plays a central role in the regulation of the Wnt and Hedgehog pathways. These pathways are critical for controlling cell fate decisions and developmental processes. In the Wnt pathway EED in conjunction with other PRC2 members modulates β-catenin-mediated transcription. In the Hedgehog pathway EED impacts the Gli family of transcription factors underlining its importance in development and cellular proliferation regulation.
EED has been linked to various cancers and developmental disorders. Aberrant EED function and localization relate to overactivation of the Wnt pathway an aspect seen in colorectal cancer. EED also interacts with EHZ2 contributing to tumorigenesis through unchecked cell division and proliferation. In the context of Weaver syndrome mutations within EED are involved consolidating its important association with developmental anomalies.
We have tested this species and application combination and it works. It is covered by our product promise.
We have not tested this specific species and application combination in-house, but expect it will work. It is covered by our product promise.
This species and application combination has not been tested, but we predict it will work based on strong homology. However, this combination is not covered by our product promise.
We do not recommend this combination. It is not covered by our product promise.
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Full details and terms and conditions can be found here:
Terms & Conditions.
Chromatin was prepared from NT2/D1 cells according to the Abcam Dual-X-ChIP protocol. Cells were fixed with 1.5 mM EGS for 30mins and then formaldehyde for 10min.
The ChIP was performed with 25 μg of chromatin, 5 μg of ab240650 (red), or 5 μg of rabbit normal IgG Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730 (gray) and 20 μl of Protein A/G sepharose beads. The immunoprecipitated DNA was quantified by real time PCR (Taqman approach for active and inactive loci, Sybr green approach for heterochromatic loci).
Primers and probes are commercial primers from Millipore (Cat. No.: 17-10034) and CST (85322S)
EED was immunoprecipitated from 0.35 mg K562 (human chronic myelogenous leukemia lymphoblast) whole cell lysate with ab240650 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab240650 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366) was used at 1/5000 dilution.
Lane 1: K562 (human chronic myelogenous leukemia lymphoblast) whole cell lysate 10 μg
Lane 2: ab240650 IP in K562 whole cell lysate
Lane 3: Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of ab240650 in K562 whole cell lysate
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 1 min
All lanes: Immunoprecipitation - Anti-EED antibody [EPR23043-5] - ChIP Grade (ab240650)
Predicted band size: 50 kDa
Blocking and dilution buffer and concentration: 5% NFDM/TBST
ab240650 detects an unknown band close to the target bands in cytoplasm.
All lanes: Western blot - Anti-EED antibody [EPR23043-5] - ChIP Grade (ab240650) at 1/1000 dilution
Lane 1: Wild type HAP1 whole cell lysate at 20 µg
Lane 2: EED knockout HAP1 whole cell lysate at 20 µg
Lane 3: Wild type HAP1 nuclear fraction lysate at 20 µg
Lane 4: EED knockout HAP1 nuclear fraction lysate at 20 µg
Lane 5: Wild type HAP1 cytoplasmic lysate at 20 µg
Lane 6: EED knockout HAP1 cytoplasmic lysate at 20 µg
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution
Predicted band size: 50 kDa
Observed band size: 50-70 kDa
Exposure time: 3s
This data was developed using ab240650, the same antibody clone in a different buffer formulation.
Blocking and dilution buffer and concentration: 5% NFDM/TBST
ab240650 detects an unknown band close to the target bands in cytoplasm.
ChIP sequencing analysis of chromatin from Mouse Embryonic Stem Cells with ab240650 at 0.1 μL/μg chromatin. Cross linking was performed for 10 minutes with 1% PFA. Primary incubation was for 16 hours at 4°C in a dilution buffer containing 20mM Tris at pH8, 1.1mM EDTA, 1.1% triton, and 167mM NaCl.
Blocking and diluting buffer and concentration: 5% NFDM/TBST ab240650 was shown to specifically react with EED in wild-type HAP1 cells as signal was lost in EED knockout cells. Wild-type and EED knockout samples were subjected to SDS-PAGE. ab240650 and Anti-GAPDH antibody [EPR16891] - Loading Control ab181602 (Rabbit anti-GAPDH loading control) were incubated 1 hour at room temperature at 1/1000 dilution and 1/200,000 dilution respectively. Blots were developed with Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (Goat Anti-Rabbit IgG H&L (HRP) ab97051) secondary antibody at 1/100,000 dilution for 1 hour at room temperature before imaging. The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID:27578866, 9584199). EED cDNA encodes 441-aa-long protein and 535-aa-long protein.
Exposure time: Lanes 1-3: 15 seconds Lanes 4-6: 37 seconds
All lanes: Western blot - Anti-EED antibody [EPR23043-5] - ChIP Grade (ab240650) at 1/1000 dilution
Lane 1: Wild type HAP1 whole cell lysate at 20 µg
Lane 2: EED knockout HAP1 whole cell lysate at 20 µg
Lane 3: K562 (human chronic myelogenous leukemia lymphoblast), whole cell lysate at 20 µg
Lane 4: 293T (human embryonic kidney epithelial cell), whole cell lysate at 20 µg
Lane 5: NIH/3T3 (mouse embryonic fibroblast), whole cell lysate at 20 µg
Lane 6: C2C12 (mouse myoblasts myoblast), whole cell lysate at 20 µg
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution
Predicted band size: 50 kDa
Observed band size: 50-70 kDa
Blocking and diluting buffer and concentration: 5% NFDM/TBST ab240650 was shown to specifically react with EED in wild-type HAP1 cells as signal was lost in EED knockout cells. Wild-type and EED knockout samples were subjected to SDS-PAGE. ab240650 and Anti-GAPDH antibody [EPR16891] - Loading Control ab181602 (Rabbit anti-GAPDH loading control) were incubated 1 hour at room temperature at 1/1000 dilution and 1/200,000 dilution respectively. Blots were developed with Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (Goat Anti-Rabbit IgG H&L (HRP) ab97051) secondary antibody at 1/100,000 dilution for 1 hour at room temperature before imaging. The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID:27578866, 9584199). EED cDNA encodes 441-aa-long protein and 535-aa-long protein.
Exposure time: Lanes 1-3: 15 seconds Lanes 4-6: 37 seconds
This data was developed using the same antibody clone in a different buffer formulation containing PBS, glycerol, azide and BSA (ab240650).
Additional bands at: 100 kDa, 70 kDa and 55 kDa in the cytoplasmic fraction (all possible non-specific binding)
This blot was produced using a 4-12% Bis-tris gel under reducing denaturing conditions. Following transfer, the membrane was blocked for 30 minutes at room temperature using 5% Milk before being incubated with ab240650 for 16 hours at 4°C.
All lanes: Western blot - Anti-EED antibody [EPR23043-5] - ChIP Grade (ab240650) at 1/1000 dilution
Lane 1: Wild type Mouse Embryonic Stem Cell (mESC) cytoplasmic extract
Lane 2: EED knockout (KO1) mESC cytoplasmic extract
Lane 3: EED knockout (KO2) mESC cytoplasmic extract
Lane 4: Wild type mESC nucleoplasm extract
Lane 5: EED knockout (KO1) mESC nucleoplasm extract
Lane 6: EED knockout (KO2) mESC nucleoplasm extract
Lane 7: Wild type mESC chromatin extract
Lane 8: EED knockout (KO1) mESC chromatin extract
Lane 9: EED knockout (KO2) mESC chromatin extract
All lanes: Goat anti-rabbit antibody conjugated to HRP at 1/5000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 50 kDa
Observed band size: 40 kDa, 47 kDa, 55 kDa, 60 kDa, 65 kDa
Exposure time: 30s
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