人EGFR In-Cell ELISA试剂盒(ab126419)
Key features and details
- Assay type: Cell-based (qualitative)
- Detection method: Colorimetric
- Sample type: Adherent cells
- Reacts with: Human
概述
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产品名称
人EGFR In-Cell ELISA试剂盒
参阅全部 EGFR 试剂盒 -
检测方法
Colorimetric -
样品类型
Adherent cells -
检测类型
Cell-based (qualitative) -
检测时间
5h 10m -
实验步骤
Multiple steps standard assay -
种属反应性
与反应: Human -
产品概述
ab126419 is a very rapid, convenient and sensitive assay kit that can monitor the activation or function of important biological pathways in cells. It can be used for measuring the relative amount of EGFR (activated) phosphorylation and screening the effects of various treatments, inhibitors (such as siRNA or chemicals), or activators in cultured Human cell lines. By determining EGFR protein phosphorylation in your experimental model system, you can verify pathway activation in your cell lines without spending excess time and effort in preparing cell lysate and performing an analysis of Western Blot.
In the EGFR Human In-Cell ELISA kit, cells are seeded into a 96 well tissue culture plate. The cells are fixed after various treatments, inhibitors or activators. After blocking, Anti-Phospho-EGFR (activated) or Anti-EGFR (primary antibody) is pipetted into the wells and incubated. The wells are washed, and HRP-conjugated anti mouse IgG (secondary antibody) is added to the wells. The wells are washed again, a TMB substrate solution is added to the wells and color develops in proportion to the amount of protein. 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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经测试应用
适用于: 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 HRP-conjugated Anti-Mouse IgG Concentrate 1 x 10µl Blocking Buffer Concentrate (5X) 1 x 20ml Fixing Solution 1 x 30ml Uncoated 96-well Microplate 1 unit Mouse Anti-EGFR Concentrate (Item H) 1 x 7µl Mouse Anti-Phospho-EGFR (active) Concentrate (Item G) 1 x 7µl Quenching Buffer Concentrate (30x) 1 x 2ml Stop Solution 1 x 14ml TMB One-Step Substrate Reagent 1 x 12ml Wash Buffer A Concentrate (20X) 1 x 30ml Wash Buffer B Concentrate (20X) 1 x 30ml -
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ab126419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 |
|---|---|---|
| ELISA |
Use at an assay dependent concentration.
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| 说明 |
|---|
|
ELISA
Use at an assay dependent concentration. |
图片
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In-Cell ELISA - EGFR Human In-Cell ELISA Kit (ab126419)
A431 cells were stimulated by different concentrations of EGF for 20 min at 37°C.
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Western blot - EGFR Human In-Cell ELISA Kit (ab126419)Western blot analysis of extracts from 100 ng/ml hEGF treated A431 cells. Phospho-EGFR (activated) and EGFR antibodies were used in both detection assays.
数据表及文件
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SDS download
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Datasheet download
文献 (3)
ab126419 被引用在 3 文献中.
- Alkahtani HM et al. Synthesis, anticancer, apoptosis-inducing activities and EGFR and VEGFR2 assay mechanistic studies of 5,5-diphenylimidazolidine-2,4-dione derivatives: Molecular docking studies. Saudi Pharm J 27:682-693 (2019). PubMed: 31297023
- El-Husseiny WM et al. Synthesis, antitumour and antioxidant activities of novel a,ß-unsaturated ketones and related heterocyclic analogues: EGFR inhibition and molecular modelling study. J Enzyme Inhib Med Chem 33:507-518 (2018). PubMed: 29455554
- Kimura I et al. Loss of epidermal growth factor receptor expression in oral squamous cell carcinoma is associated with invasiveness and epithelial-mesenchymal transition. Oncol Lett 11:201-207 (2016). PubMed: 26870189