人EGFR (pY1086) + total EGFR ELISA试剂盒(ab126441)
Key features and details
- Sample type: Cell Lysate
- Detection method: Colorimetric
- Assay type: Sandwich (qualitative)
- Reacts with: Human
概述
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产品名称
人EGFR (pY1086) + total EGFR ELISA试剂盒
参阅全部 EGFR 试剂盒 -
检测方法
Colorimetric -
样品类型
Cell Lysate -
检测类型
Sandwich (qualitative) -
检测时间
5h 00m -
实验步骤
Multiple steps standard assay -
种属反应性
与反应: Human -
产品概述
ab126441 is a very rapid, convenient and sensitive assay kit that can monitor the activation or function of important biological pathways in cell lysates. By determining phosphorylated EGFR protein in your experimental model system, you can verify pathway activation in your cell lysates. You can simultaneously measure numerous different cell lysates without spending excess time and effort in performing a Western Blotting analysis.
This sandwich ELISA kit is an in vitro enzyme-linked immunosorbent assay for the measurement of human phospho-EGFR (Y1086) and total EGFR (help normalize the results of phospho-EGFR from different cell lysates being compared). An anti-EGFR antibody has been coated onto a 96-well plate. Samples are pipetted into the wells and EGFR present in a sample is bound to the wells by the immobilized antibody. The wells are washed and anti-phosphorylated EGFR (Y1086) or anti-total EGFR antibody is used to detect phosphorylated or non-phosphorylated EGFR. After washing away unbound antibody, HRP-conjugated anti-Rabbit IgG or HRP-Streptavidin is pipetted to the wells. The wells are again washed, a TMB substrate solution is added to the wells and color develops in proportion to the amount of EGFR (Y1086) or total EGFR bound. The Stop Solution changes the color from blue to yellow, and the intensity of the color is measured at 450 nm.
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说明
Abcam has not and does not intend to apply for the REACH Authorisation of customers’ uses of products that contain European Authorisation list (Annex XIV) substances.
It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses. -
经测试应用
适用于: Sandwich ELISAmore details -
平台
Microplate
性能
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存放说明
Store at -20°C. Please refer to protocols. -
组件 1 x 96 tests 20X Wash Buffer 1 x 25ml 2X Cell Lysis Buffer 1 x 5ml 500X HRP-conjugated anti-rabbit IgG 1 x 25µl 5X Assay Diluent 1 x 15ml 600X HRP-Streptavidin Concentrate 1 x 200µl Anti-phospho-EGFR (Y1086) 1 vial Biotinylated-anti-EGFR 1 vial EGFR Microplate (12 strips x 8 wells) coated with monoclonal anti-EGFR antibody 1 unit Positive Control: lyophilized powder from A431 cell lysate 1 vial Stop Solution 1 x 8ml TMB One-Step Substrate Reagent 1 x 12ml -
研究领域
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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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别名
- Avian erythroblastic leukemia viral (v erb b) oncogene homolog
- Cell growth inhibiting protein 40
- Cell proliferation inducing protein 61
see all -
数据库链接
- Entrez Gene: 1956 Human
- Omim: 131550 Human
- SwissProt: P00533 Human
- Unigene: 488293 Human
- Unigene: 605083 Human
应用
The Abpromise guarantee
Abpromise™承诺保证使用ab126441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 |
|---|---|---|
| Sandwich ELISA |
Use at an assay dependent concentration.
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| 说明 |
|---|
|
Sandwich ELISA
Use at an assay dependent concentration. |
图片
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Sandwich ELISA - EGFR (pY1086) + total EGFR Human ELISA Kit (ab126441)A431 cells were treated with recombinant human EGF at 37°C for 20 min. Solubilize cells at 4 x 107 cells/ml in Cell Lysate Buffer. Serial dilutions of lysates were analyzed in this ELISA.
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Western blot - EGFR (pY1086) + total EGFR ELISA Kit (ab126441)The A431 cells were treated with 100 ng/ml recombinant human EGF for 20 minutes to induce phosphorylation of EGFR. Serial dilutions of lysates were analyzed by Western blot. Immunoblots were incubated with anti-phospho-EGFR (Y1086). -
Sandwich ELISA - EGFR (pY1086) + total EGFR ELISA Kit (ab126441)The A431 cells were treated with 100 ng/ml recombinant human EGF for 20 minutes to induce phosphorylation of EGFR. Serial dilutions of lysates were analyzed in this ELISA. -
Western blot - EGFR (pY1086) + total EGFR ELISA Kit (ab126441)A431 cells were treated or untreated with 100 ng/ml recombinant human EGF for 10 min. Cell lysates were analyzed by Western Blot -
Sandwich ELISA - EGFR (pY1086) + total EGFR ELISA Kit (ab126441)A431 cells were treated or untreated with 100 ng/ml recombinant human EGF for 10 min. Cell lysates were analyzed using this phosphoELISA.
数据表及文件
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SDS download
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Datasheet download
文献 (1)
ab126441 被引用在 1 文献中.
- Zhi Y et al. Synthesis and pharmacological evaluation of novel epidermal growth factor receptor inhibitors against prostate tumor cells. Oncol Lett 16:6522-6530 (2018). PubMed: 30405791