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Epigenetics and Nuclear Signaling Chromatin Modifying Enzymes Methylation
使用敲除细胞株进行验证

Anti-EED抗体(ab4469)

  • Datasheet
  • SDS
Reviews (3)Q&A (1)References (29)

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Western blot - Anti-EED antibody (ab4469)
  • Immunocytochemistry/ Immunofluorescence - Anti-EED antibody (ab4469)
  • Western blot - 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

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概述

  • 产品名称

    Anti-EED抗体
    参阅全部 EED 一抗
  • 描述

    兔多克隆抗体to EED
  • 宿主

    Rabbit
  • 经测试应用

    适用于: WB, ICC/IFmore details
  • 种属反应性

    与反应: Human, Recombinant fragment
    预测可用于: Mouse, Chicken, Cow, Xenopus laevis
  • 免疫原

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

  • 阳性对照

    • WB: Human Recombinant EED Protein, HAP1, HeLa and HepG2 cell lysates ICC/IF: U-2 OS cells
  • 常规说明

    The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.

    If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As

性能

  • 形式

    Liquid
  • 存放说明

    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.
  • 存储溶液

    pH: 7.40
    Preservative: 0.02% Sodium azide
    Constituent: PBS

    Batches of this product that have a concentration < 1mg/ml may have BSA added as a stabilising agent. If you would like information about the formulation of a specific lot, please contact our scientific support team who will be happy to help.
  • Concentration information loading...
  • 纯度

    Immunogen affinity purified
  • 克隆

    多克隆
  • 同种型

    IgG
  • 研究领域

    • Epigenetics and Nuclear Signaling
    • Chromatin Modifying Enzymes
    • Methylation
    • Epigenetics and Nuclear Signaling
    • Chromatin Remodeling
    • Polycomb Silencing
    • PRC2
    • Epigenetics and Nuclear Signaling
    • Chromatin Remodeling
    • Polycomb Silencing
    • PRC3
    • Epigenetics and Nuclear Signaling
    • Chromatin Modifying Enzymes
    • Methylation
    • Lysine methylation

相关产品

  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
    • Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773)
  • Isotype control

    • Rabbit IgG, polyclonal - Isotype Control (ChIP Grade) (ab171870)
  • Recombinant Protein

    • Recombinant Human EED protein (ab156723)

应用

The Abpromise guarantee

Abpromise™承诺保证使用ab4469于以下的经测试应用

“应用说明”部分 下显示的仅为推荐的起始稀释度;实际最佳的稀释度/浓度应由使用者检定。

应用 Ab评论 说明
WB (3)
1/200. Predicted molecular weight: 49 kDa.
ICC/IF
1/100.
说明
WB
1/200. Predicted molecular weight: 49 kDa.
ICC/IF
1/100.

靶标

  • 功能

    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.
  • 组织特异性

    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.
  • 序列相似性

    Belongs to the WD repeat ESC family.
    Contains 7 WD repeats.
  • 发展阶段

    Expression peaks at the G1/S phase boundary.
  • 细胞定位

    Nucleus. Chromosome. Transiently colocalizes with XIST at inactive X chromosomes.
  • Target information above from: UniProt accession O75530 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • 数据库链接

    • 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
    • SwissProt: O75530 Human
    • SwissProt: Q921E6 Mouse
    • SwissProt: Q8UUP2 Xenopus laevis
    • Unigene: 503510 Human
    • Unigene: 380914 Mouse
    see all
  • 别名

    • eed antibody
    • EED protein antibody
    • EED, mouse, homolog of antibody
    • EED_HUMAN antibody
    • Embryonic ectoderm development antibody
    • Embryonic ectoderm development isoform a antibody
    • Embryonic ECTODERM development protein homolog antibody
    • Embryonic ectoderm development protein, mouse, homolog of antibody
    • hEED antibody
    • OTTHUMP00000235483 antibody
    • OTTHUMP00000235484 antibody
    • OTTHUMP00000235485 antibody
    • Polycomb protein eed antibody
    • WAIT 1 antibody
    • WAIT-1 antibody
    • WAIT1 antibody
    • WD protein associating with integrin cytoplasmic tails 1 antibody
    see all

图片

  • Western blot - Anti-EED antibody (ab4469)
    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)
    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)
    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).

实验方案

  • Western blot protocols
  • Immunocytochemistry & immunofluorescence protocols

Click here to view the general protocols

数据表及文件

  • SDS download

  • Datasheet download

    Download

文献 (29)

发表研究结果有使用 ab4469?请让我们知道,以便我们可以引用本数据表中的参考文章。

ab4469 被引用在 29 文献中.

  • Boulay G  et al. The chromatin landscape of primary synovial sarcoma organoids is linked to specific epigenetic mechanisms and dependencies. Life Sci Alliance 4:N/A (2021). PubMed: 33361335
  • Ngan CY  et al. Chromatin interaction analyses elucidate the roles of PRC2-bound silencers in mouse development. Nat Genet 52:264-272 (2020). PubMed: 32094912
  • Ma P  et al. RNF220 is required for cerebellum development and regulates medulloblastoma progression through epigenetic modulation of Shh signaling. Development 147:N/A (2020). PubMed: 32376680
  • Wei D  et al. Novel renal medullary carcinoma cell lines, UOK353 and UOK360, provide preclinical tools to identify new therapeutic treatments. Genes Chromosomes Cancer 59:472-483 (2020). PubMed: 32259323
  • Tan X  et al. Inhibition of EZH2 enhances the therapeutic effect of 5-FU via PUMA upregulation in colorectal cancer. Cell Death Dis 11:1061 (2020). PubMed: 33311453
View all Publications for this product

客户评价及客户问答

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1-4 of 4 Abreviews or Q&A

Western blot abreview for Anti-EED antibody

Excellent
Abreviews
Abreviews
abreview image
Application
Western blot
Sample
Human Cell lysate - whole cell (THP1)
Gel Running Conditions
Reduced Denaturing (10% SDS-PAGE)
Loading amount
30 µg
Specification
THP1
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

提交于 Sep 06 2017

Western blot abreview for Anti-EED antibody

Good
Abreviews
Abreviews
abreview image
Application
Western blot
Sample
Human Cell lysate - whole cell (DLD1)
Gel Running Conditions
Reduced Denaturing (10%)
Loading amount
20 µg
Specification
DLD1
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

提交于 Feb 02 2017

Western blot abreview for Anti-EED antibody

Inconclusive
Abreviews
Abreviews
abreview image
Application
Western blot
Loading amount
30 µg
Gel Running Conditions
Non-reduced Denaturing (10%)
Sample
Mouse Cell lysate - whole cell (Extra Embryonic Endoderm Cells (XEN))
Specification
Extra Embryonic Endoderm Cells (XEN)
Blocking step
BSA as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 25°C
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

提交于 Nov 27 2013

Question

I had tried 5 times used the ab4469 antibody and It did not work. And I had tried 5 times, so I bought another antibody UPSTATE:17-663, and had got the anticipate result. The condition were as follows: The concentration of the primary antibody was 1:1000; The concentration of the secondary antibody was 1:5000;   So I thought there was something wrong with this antibody and did not want to try again.  

Read More

Abcam community

Verified customer

Asked on Sep 07 2011

Answer

Thank you for contacting us. I am sorry that the results did not improve. I would like to ask how you would like to proceed with this case. In order to help you further I would need more information about order number. The order number you have provided does not match actual Abcam order. It looks like you are from China and you may have purchased this antibody from our Chinese distributor. Please contact them for contacting us for further instructions. Please be advised that I can only provide free of charge replacement or refund/ credit note if the antibody was ordered within Abcam guarantee period; which is 4 month for Chinese customers. If you are not from China please accept my apologies for misunderstanding and provide me the correct 6 digit Abcam order number. I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

Read More

Abcam Scientific Support

回复于 Sep 07 2011

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
For licensing inquiries, please contact partnerships@abcam.com

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