Human EGFR In-Cell ELISA Kit (ab126419)
- Datasheet
- References (1)
- Protocols
Overview
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Product nameHuman EGFR In-Cell ELISA Kit
See all EGFR kits -
Detection methodColorimetric
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Sample typeAdherent cells
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Assay typeCell-based (qualitative)
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Assay time5h 10m
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Assay durationMultiple steps standard assay
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Species reactivityReacts with: Human
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Product overview
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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Tested applicationsSuitable for: In-Cell ELISAmore details
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PlatformMicroplate
Properties
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Storage instructionsStore at -20°C. Please refer to protocols.
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Components 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 -
Research areas
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FunctionReceptor for EGF, but also for other members of the EGF family, as TGF-alpha, amphiregulin, betacellulin, heparin-binding EGF-like growth factor, GP30 and vaccinia virus growth factor. Is involved in the control of cell growth and differentiation. Phosphorylates MUC1 in breast cancer cells and increases the interaction of MUC1 with SRC and CTNNB1/beta-catenin.
Isoform 2 may act as an antagonist of EGF action. -
Tissue specificityUbiquitously expressed. Isoform 2 is also expressed in ovarian cancers.
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Involvement in diseaseDefects in EGFR are associated with lung cancer (LNCR) [MIM:211980].
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Sequence similaritiesBelongs to the protein kinase superfamily. Tyr protein kinase family. EGF receptor subfamily.
Contains 1 protein kinase domain. -
Post-translational
modificationsPhosphorylation of Ser-695 is partial and occurs only if Thr-693 is phosphorylated.
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 occur. -
Cellular localizationSecreted and Cell membrane. Endoplasmic reticulum membrane. Golgi apparatus membrane. Nucleus membrane. Endosome. In response to EGF, translocated from the cell membrane to the nucleus via Golgi and ER. Co-localizes with TNK2 on the endosomes.
- Information by UniProt
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Alternative names
- 9030024J15Rik
- AI552599
- Avian erythroblastic leukemia viral (v erb b) oncogene homolog
see all -
Database links
- Entrez Gene: 1956 Human
- Omim: 131550 Human
- SwissProt: P00533 Human
- Unigene: 488293 Human
Applications
Our Abpromise guarantee covers the use of ab126419 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
| Application | Abreviews | Notes |
|---|---|---|
| In-Cell ELISA | Use at an assay dependent concentration. |
Images
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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. -
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.
Protocols
References
This product has been referenced in:
- 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). Read more (PubMed: 29455554) »
