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Rabbit Recombinant Monoclonal EED antibody. Suitable for IP, ChIP, WB, ChIP-seq and reacts with Human, Mouse samples. Cited in 2 publications.


Images

ChIP - Anti-EED antibody [EPR23043-5] - ChIP Grade (AB240650), expandable thumbnail
  • Immunoprecipitation - Anti-EED antibody [EPR23043-5] - ChIP Grade (AB240650), expandable thumbnail
  • Western blot - Anti-EED antibody [EPR23043-5] - ChIP Grade (AB240650), expandable thumbnail
  • ChIP-sequencing - Anti-EED antibody [EPR23043-5] - ChIP Grade (AB240650), expandable thumbnail
  • Western blot - Anti-EED antibody [EPR23043-5] - ChIP Grade (AB240650), expandable thumbnail

Publications

Key facts

Isotype

IgG

Host species

Rabbit

Storage buffer

Preservative: 0.01% Sodium azide
Constituents: 59.94% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA

Form

Liquid

Clonality

Monoclonal

Immunogen

  • The exact immunogen used to generate this antibody is proprietary information.

Reactivity data

Select an application
Product promiseTestedExpectedPredictedNot recommended
IPChIPWBIHC-PICC/IFChIP-seqFlow 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

Tested
Tested

Species

Human

Dilution info

1/30

Notes

-

Expected
Expected

Species

Mouse

Dilution info

Use at an assay dependent concentration.

Notes

-

Not recommended
Not recommended

Species

Rat

Dilution info

-

Notes

-

Tested
Tested

Species

Human

Dilution info

5 µg chromatin for 25.00000 µg chromatin

Notes

-

Expected
Expected

Species

Mouse

Dilution info

Use at an assay dependent concentration.

Notes

-

Not recommended
Not recommended

Species

Rat

Dilution info

-

Notes

-

Tested
Tested

Species

Mouse

Dilution info

1/1000

Notes

-

Species

Human

Dilution info

1/1000

Notes

-

Not recommended
Not recommended

Species

Rat

Dilution info

-

Notes

-

Not recommended
Not recommended

Species

Mouse, Human, Rat

Dilution info

-

Notes

-

Not recommended
Not recommended

Species

Mouse, Human, Rat

Dilution info

-

Notes

-

Tested
Tested

Species

Mouse

Dilution info

0.1 µL for 1.00000 µg chromatin

Notes

Use at 0.1 uL/ug chromatin.

Expected
Expected

Species

Human

Dilution info

Use at an assay dependent concentration.

Notes

-

Not recommended
Not recommended

Species

Rat

Dilution info

-

Notes

-

Not recommended
Not recommended

Species

Human, Mouse, Rat

Dilution info

-

Notes

-

Target data

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. 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.

Alternative names

Recommended products

Rabbit Recombinant Monoclonal EED antibody. Suitable for IP, ChIP, WB, ChIP-seq and reacts with Human, Mouse samples. Cited in 2 publications.

Key facts

Isotype

IgG

Form

Liquid

Clonality

Monoclonal

Immunogen
  • The exact immunogen used to generate this antibody is proprietary information.
Clone number

EPR23043-5

Purification technique

Affinity purification Protein A

Specificity

ab240650 detects an unknown band close to the target bands in cytoplasm.

Concentration
Loading...

Storage

Shipped at conditions

Blue Ice

Appropriate short-term storage duration

1-2 weeks

Appropriate short-term storage conditions

+4°C

Appropriate long-term storage conditions

-20°C

Aliquoting information

Upon delivery aliquot

Storage information

Avoid freeze / thaw cycle

Notes

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:

  • - High batch-to-batch consistency and reproducibility
  • - Improved sensitivity and specificity
  • - Long-term security of supply
  • - Animal-free batch production

For more information, read more on recombinant antibodies.

Supplementary info

This supplementary information is collated from multiple sources and compiled automatically.

Activity summary

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.

Biological function summary

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.

Pathways

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.

Associated diseases and disorders

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.

Product promise

We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.

In the unlikely event of one of our products not working as expected, you are covered by our product promise.

Full details and terms and conditions can be found here:
Terms & Conditions.

6 product images

  • ChIP - Anti-EED antibody [EPR23043-5] - ChIP Grade (ab240650), expandable thumbnail

    ChIP - Anti-EED antibody [EPR23043-5] - ChIP Grade (ab240650)

    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)

  • Immunoprecipitation - Anti-EED antibody [EPR23043-5] - ChIP Grade (ab240650), expandable thumbnail

    Immunoprecipitation - Anti-EED antibody [EPR23043-5] - ChIP Grade (ab240650)

    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

  • Western blot - Anti-EED antibody [EPR23043-5] - ChIP Grade (ab240650), expandable thumbnail

    Western blot - Anti-EED antibody [EPR23043-5] - ChIP Grade (ab240650)

    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

    Secondary

    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 - Anti-EED antibody [EPR23043-5] - ChIP Grade (ab240650), expandable thumbnail
    This image is courtesy of a customer review submitted by Ivano Mocavini.

    ChIP-sequencing - Anti-EED antibody [EPR23043-5] - ChIP Grade (ab240650)

    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.

  • Western blot - Anti-EED antibody [EPR23043-5] - ChIP Grade (ab240650), expandable thumbnail

    Western blot - Anti-EED antibody [EPR23043-5] - ChIP Grade (ab240650)

    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

    Secondary

    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).

  • Western blot - Anti-EED antibody [EPR23043-5] - ChIP Grade (ab240650), expandable thumbnail
    This image is courtesy of a customer review submitted by Ivano Mocavini.

    Western blot - Anti-EED antibody [EPR23043-5] - ChIP Grade (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

    Secondary

    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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Product protocols

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