重组Anti-EGFR (phospho S1046 + S1047)抗体[EP2259Y] (ab76300)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EP2259Y] to EGFR (phospho S1046 + S1047)
- Suitable for: WB, IP, ICC/IF, Dot blot
- Reacts with: Human
Related conjugates and formulations
概述
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产品名称
Anti-EGFR (phospho S1046 + S1047)抗体[EP2259Y]
参阅全部 EGFR 一抗 -
描述
兔单克隆抗体[EP2259Y] to EGFR (phospho S1046 + S1047) -
宿主
Rabbit -
特异性
Recognises EGFR phosphorylated on Serine 1046 and Serine 1047 of the mature human isoform 1 (corresponding to S1070 and S1071 from the precursor form P00533-1/p170) -
经测试应用
适用于: WB, IP, ICC/IF, Dot blotmore details
不适用于: Flow Cyt or IHC-P -
种属反应性
与反应: 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: A431 cell lysates treated with EGF. ICC/IF: A431 cells treated with EGF. IP: A431 treated with EGF whole cell lysate. Dot Blot: EGFR (pS1046/pS1047) phospho peptide.
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常规说明
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.
性能
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形式
Liquid -
存放说明
Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles. -
存储溶液
pH: 7.20
Preservative: 0.05% Sodium azide
Constituents: 0.1% BSA, 40% Glycerol (glycerin, glycerine), 9.85% Tris glycine, 50% Tissue culture supernatant -
Concentration information loading...
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纯度
Protein A purified -
克隆
单克隆 -
克隆编号
EP2259Y -
同种型
IgG -
研究领域
相关产品
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Alternative Versions
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Isotype control
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Recombinant Protein
应用
The Abpromise guarantee
Abpromise™承诺保证使用ab76300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 |
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WB | (1) |
1/500000 - 1/1e+006. Detects a band of approximately 150 kDa (predicted molecular weight: 134 kDa).
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IP |
1/40.
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ICC/IF |
1/100 - 1/250.
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Dot blot |
1/1000.
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说明 |
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WB
1/500000 - 1/1e+006. Detects a band of approximately 150 kDa (predicted molecular weight: 134 kDa). |
IP
1/40. |
ICC/IF
1/100 - 1/250. |
Dot blot
1/1000. |
靶标
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功能
Receptor tyrosine kinase binding ligands of the EGF family and activating several signaling cascades to convert extracellular cues into appropriate cellular responses. Known ligands include EGF, TGFA/TGF-alpha, amphiregulin, epigen/EPGN, BTC/betacellulin, epiregulin/EREG and HBEGF/heparin-binding EGF. Ligand binding triggers receptor homo- and/or heterodimerization and autophosphorylation on key cytoplasmic residues. The phosphorylated receptor recruits adapter proteins like GRB2 which in turn activates complex downstream signaling cascades. Activates at least 4 major downstream signaling cascades including the RAS-RAF-MEK-ERK, PI3 kinase-AKT, PLCgamma-PKC and STATs modules. May also activate the NF-kappa-B signaling cascade. Also directly phosphorylates other proteins like RGS16, activating its GTPase activity and probably coupling the EGF receptor signaling to the G protein-coupled receptor signaling. Also phosphorylates MUC1 and increases its interaction with SRC and CTNNB1/beta-catenin.
Isoform 2 may act as an antagonist of EGF action. -
组织特异性
Ubiquitously expressed. Isoform 2 is also expressed in ovarian cancers. -
疾病相关
Lung cancer
Inflammatory skin and bowel disease, neonatal, 2 -
序列相似性
Belongs to the protein kinase superfamily. Tyr protein kinase family. EGF receptor subfamily.
Contains 1 protein kinase domain. -
翻译后修饰
Phosphorylation at Ser-695 is partial and occurs only if Thr-693 is phosphorylated. Phosphorylation at Thr-678 and Thr-693 by PRKD1 inhibits EGF-induced MAPK8/JNK1 activation. Dephosphorylation by PTPRJ prevents endocytosis and stabilizes the receptor at the plasma membrane. Autophosphorylation at Tyr-1197 is stimulated by methylation at Arg-1199 and enhances interaction with PTPN6. Autophosphorylation at Tyr-1092 and/or Tyr-1110 recruits STAT3. Dephosphorylated by PTPN1 and PTPN2.
Monoubiquitinated and polyubiquitinated upon EGF stimulation; which does not affect tyrosine kinase activity or signaling capacity but may play a role in lysosomal targeting. Polyubiquitin linkage is mainly through 'Lys-63', but linkage through 'Lys-48', 'Lys-11' and 'Lys-29' also occurs. Deubiquitination by OTUD7B prevents degradation. Ubiquitinated by RNF115 and RNF126.
Methylated. Methylation at Arg-1199 by PRMT5 stimulates phosphorylation at Tyr-1197. -
细胞定位
Secreted and Cell membrane. Endoplasmic reticulum membrane. Golgi apparatus membrane. Nucleus membrane. Endosome. Endosome membrane. Nucleus. In response to EGF, translocated from the cell membrane to the nucleus via Golgi and ER. Endocytosed upon activation by ligand. Colocalized with GPER1 in the nucleus of estrogen agonist-induced cancer-associated fibroblasts (CAF). - Information by UniProt
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数据库链接
- Entrez Gene: 1956 Human
- Omim: 131550 Human
- SwissProt: P00533 Human
- Unigene: 488293 Human
- Unigene: 605083 Human
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别名
- Avian erythroblastic leukemia viral (v erb b) oncogene homolog antibody
- Cell growth inhibiting protein 40 antibody
- Cell proliferation inducing protein 61 antibody
see all
图片
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All lanes : Anti-EGFR (phospho S1046 + S1047) antibody [EP2259Y] (ab76300) at 1/1000000 dilution
Lane 1 : A431 cell lysate
Lane 2 : A431 cell lysate treated with EGF
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : HRP labelled goat anti-rabbit at 1/2000 dilution
Predicted band size: 134 kDa
Observed band size: 180 kDa why is the actual band size different from the predicted? -
Immunocytochemistry/Immunofluorescence analysis of untreated and EGF (100ng/mL for 10 minutes) treated A431 cells labelling EGFR (phospho S1046 + S1047) with ab76300 at 1/100. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody. The cells were co-stained with ab195889, an Alexa Fluor® 594-conjugated mouse anti-tubulin (1/200). Nuclei counterstained with DAPI (blue).
Control 1: untreated A431 cells incubated with PBS instead of primary antibody followed by incubation with ab150077 Alexa Fluor® 488 goat anti-rabbit IgG.
Control 2: EGF treated A431 cells incubated with PBS instead of primary antibody followed by incubation with ab150077 Alexa Fluor® 488 goat anti-rabbit IgG. -
Purified ab76300 at 1/40 dilution (2µg) immunoprecipitating EGFR in A431 treated with EGF whole cell lysate.
Lane 1 (input): A431 (Human epidermoid carcinoma epithelial cell) treated with EGF whole cell lysate 10µg
Lane 2 (+): ab76300 + A431 treated with EGF whole cell lysate.
Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab76300 in A431 treated with EGF whole cell lysate.
VeriBlot for IP Detection Reagent (HRP) (ab131366) (1/1000 dilution) was used for Western blotting.
Blocking Buffer and concentration: 5% NFDM/TBST.
Diluting buffer and concentration: 5% NFDM/TBST.
Observed band size: 180 kDa -
Primary antibody: ab76300 at a dilution of 1/1000.
Secondary antibody: Peroxidase conjugated-goat anti-rabbit IgG, (H+L) at a dilution of 1/2500.
Blocking and dilution buffer: 5% NFDM/TBST
Lane 1: EGFR (pS1046/pS1047) phospho peptide.
Lane 2: EGFR non-phospho peptide.Exposure time: 3 minutes.
实验方案
数据表及文件
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SDS download
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Datasheet download
文献 (1)
ab76300 被引用在 1 文献中.
- Singh RK & Lokeshwar BL The IL-8-regulated chemokine receptor CXCR7 stimulates EGFR signaling to promote prostate cancer growth. Cancer Res 71:3268-77 (2011). PubMed: 21398406