重组Anti-EED抗体[EPR23043-5] - ChIP Grade (ab240650)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR23043-5] to EED - ChIP Grade
- Suitable for: ChIP, ChIP-sequencing, WB, IP
- Knockout validated
- Reacts with: Mouse, Human
Related conjugates and formulations
概述
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产品名称
Anti-EED抗体[EPR23043-5] - ChIP Grade
参阅全部 EED 一抗 -
描述
兔单克隆抗体[EPR23043-5] to EED - ChIP Grade -
宿主
Rabbit -
特异性
ab240650 detects an unknown band close to the target bands in cytoplasm.
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经测试应用
适用于: ChIP, ChIP-sequencing, WB, IPmore details
不适用于: Flow Cyt,ICC/IF or IHC-P -
种属反应性
与反应: Mouse, Human -
免疫原
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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阳性对照
- WB: Wild type HAP1, K562, 293T, NIH/3T3 and C2C12 lysates. IP: K562 cells. ChIP: Chromatin prepared from NT2/D1 cells.
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常规说明
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 production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
性能
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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 long term. Avoid freeze / thaw cycle. -
存储溶液
Preservative: 0.01% Sodium azide
Constituents: 59.94% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading...
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纯度
Protein A purified -
克隆
单克隆 -
克隆编号
EPR23043-5 -
同种型
IgG -
研究领域
相关产品
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Alternative Versions
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Compatible Secondaries
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Related Products
应用
The Abpromise guarantee
Abpromise™承诺保证使用ab240650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 |
---|---|---|
ChIP | (3) |
Use 5 µg for 25 µg of chromatin.
|
ChIP-sequencing |
Use a concentration of 0.1 µl/chromatin.
Use at 0.1 uL/ug chromatin. |
|
WB | (2) |
1/1000. Predicted molecular weight: 50 kDa.
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IP |
1/30.
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说明 |
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ChIP
Use 5 µg for 25 µg of chromatin. |
ChIP-sequencing
Use a concentration of 0.1 µl/chromatin. Use at 0.1 uL/ug chromatin. |
WB
1/1000. Predicted molecular weight: 50 kDa. |
IP
1/30. |
靶标
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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. - Information by UniProt
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数据库链接
- Entrez Gene: 8726 Human
- Entrez Gene: 13626 Mouse
- Omim: 605984 Human
- SwissProt: O75530 Human
- SwissProt: Q921E6 Mouse
- Unigene: 503510 Human
- Unigene: 380914 Mouse
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别名
- eed antibody
- EED protein antibody
- EED, mouse, homolog of antibody
see all
图片
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All lanes : Anti-EED antibody [EPR23043-5] - ChIP Grade (ab240650) at 1/1000 dilution
Lane 1 : Wild type HAP1 whole cell lysate
Lane 2 : EED knockout HAP1 whole cell lysate
Lane 3 : Wild type HAP1 nuclear fraction lysate
Lane 4 : EED knockout HAP1 nuclear fraction lysate
Lane 5 : Wild type HAP1 cytoplasmic lysate
Lane 6 : EED knockout HAP1 cytoplasmic lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 50 kDa
Observed band size: 50-70 kDa why is the actual band size different from the predicted?
Exposure time: 3 secondsBlocking and dilution buffer and concentration: 5% NFDM/TBST
ab240650 detects an unknown band close to the target bands in cytoplasm.
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All lanes : Anti-EED antibody [EPR23043-5] - ChIP Grade (ab240650) at 1/1000 dilution
Lane 1 : Wild type HAP1 whole cell lysate
Lane 2 : EED knockout HAP1 whole cell lysate
Lane 3 : K562 (human chronic myelogenous leukemia lymphoblast), whole cell lysate
Lane 4 : 293T (human embryonic kidney epithelial cell), whole cell lysate
Lane 5 : NIH/3T3 (mouse embryonic fibroblast), whole cell lysate
Lane 6 : C2C12 (mouse myoblasts myoblast), whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 50 kDa
Observed band size: 50-70 kDa why is the actual band size different from the predicted?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 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 (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
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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) (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 (ab172730) instead of ab240650 in K562 whole cell lysate
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 1 min
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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 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)
*https://www.abcam.com/resources?keywords=X%20ChIP%20protocol
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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.
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All lanes : 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: 40,47,55,60,65 kDa why is the actual band size different from the predicted?
Exposure time: 30 secondsAdditional 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.
实验方案
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
数据表及文件
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SDS download
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Datasheet download
Certificate of Compliance
文献 (1)
ab240650 被引用在 1 文献中.
- Weigert R et al. Dynamic antagonism between key repressive pathways maintains the placental epigenome. Nat Cell Biol 25:579-591 (2023). PubMed: 37024684