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Multispecies EGFR In-Cell ELISA Kit, Colorimetric
Evaluation of dose-dependent activation and inhibition of EGFR in A431 cells with EGF and PD169393. Panels A and B show an increase in phospho-EGFR with increasing doses of EGF. The EGF-induced phosphorylation of EGFR is inhibited with increasing concentrations of PD169393 (panel C, rows C and D), whereas total EGFR does not significantly change with either treatment. No significant staining is seen in the absence of primary antibodies (panel A, column 1 and panel C, column 12). Panels B and D show that the fluorescence (RIU, left Y-axis) ... View More
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Product Specifications
Sample type/volume
Supernatant
10,000 cells
Hands-on time
1 hr 20 min
Time-to-result
24 hr
Homogenous (no wash)
No
Instrument
Absorbance Microplate Reader
Product size
96 Tests
Contents
Blocking Buffer
20X Tris Buffered Saline
Surfact-Amps 20
Surfact-Amps X-100
HRP Conjugate
TMB Substrate
TMB Stop Solution
Janus Green Whole-Cell Stain
Elution Buffer
Thin Plate Seal Assembly
Components included only in the target-specific kits:Antibody #1 and Antibody #2
20X Tris Buffered Saline
Surfact-Amps 20
Surfact-Amps X-100
HRP Conjugate
TMB Substrate
TMB Stop Solution
Janus Green Whole-Cell Stain
Elution Buffer
Thin Plate Seal Assembly
Components included only in the target-specific kits:Antibody #1 and Antibody #2
Shipping conditions
Wet ice
Storage
2-8°C
Protein name
EGFR
Target site-specificity
Protein family
Growth Factors & Receptors
Assay kit format
In-cell ELISA Kit
Detector antibody conjugate
HRP
Label or dye
HRP
About This Kit
The EGFR Multispecies In-Cell ELISA Colorimetric Detection Kit is a simple method for quantifying intracellular proteins in whole cells.
Principle of the method
To perform the assay, cells are first plated, treated and fixed. Expression of the protein(s) of interest is monitored in wells of a microplate using target-specific primary antibodies (see the Important Product Information section for antibodies included in each kit) and a horseradish peroxidase (HRP)-conjugated detection reagent. The kit is supplied with a whole-cell stain to control for differences in cell plating, which is important when measuring relative levels of a protein with different treatments or assessing its post-translational modification (PTM) form. After staining, the results are analyzed by normalizing the absorbance (HRP activity) values to cell number, which adjusts for the cell plating differences among the wells.
Rigorous validation
Each manufactured lot of this ELISA kit is quality tested for criteria such as sensitivity, specificity, precision, and lot-to-lot consistency. See manual for more information on validation.
Target information
EGFR, epidermal growth factor receptor, is a receptor tyrosine kinases that signals in response to various growth factors. Overexpression has been linked to numerous types of cancer and EGFR is the target of both biological and small molecular therapeutics. EGFR is encoded by the EGFR gene located on chromosome 7 in humans. EGFR belongs to the HER/ERbB family of proteins that includes three other receptor tyrosine kinases, ERbB2, ERbB3, ERbB4. EGFR is a transmembrane receptor and binding of its cognate ligands such as EGF (Epidermal Growth Factor) and TGF alpha (Transforming Growth Factor alpha) to the extracellular domain leads to EGFR dimerization followed by autophosphorylation of the tyrosine residues in the cytoplasmic domain. Overexpression is observed in tumors of the head and neck, brain, bladder, stomach, breast, lung, endometrium, cervix, vulva, ovary, esophagus, stomach and in squamous cell carcinoma.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
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Bioinformatics
Protein Aliases : Proto-oncogene c-ErbB-1, Receptor tyrosine-protein kinase erbB-1
Performance Guarantee
If an Invitrogen™ antibody doesn't perform as described on our website or datasheet,we'll replace the product at no cost to you, or provide you with a credit for a future purchase.*
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