Phospho-FGFR2 alpha DuoSet IC ELISA Summary
* Provided that the recommended microplates, buffers, diluents, substrates and solutions are used, and the assay is run as summarized in the Assay Procedure provided.
- Optimized capture and detection antibody pairings and recommended concentrations save lengthy development time
- Development protocols are provided to guide further assay optimization
- Assay can be customized to your specific needs
- Available in 2, 5, and 15-(96-well) plate pack sizes
- Economical alternative to Western blot
- Capture Antibody
- Conjugated Detection Antibody
- Calibrated Immunoassay Standard or Control
Other Reagents Required
PBS: (Catalog # DY006), or 137 mM NaCl, 2.7 mM KCl, 8.1 mM Na2HPO4, 1.5 mM KH2O4, pH 7.2 - 7.4, 0.2 µm filtered
Wash Buffer: (Catalog # WA126), or equivalent
Substrate Solution: 1:1 mixture of Color Reagent A (H2O2) and Color Reagent B (Tetramethylbenzidine) (Catalog # DY999)
Stop Solution: 2 N H2SO4 (Catalog # DY994)
Microplates: R&D Systems (Catalog # DY990), or equivalent
Plate Sealers: ELISA Plate Sealers (Catalog # DY992), or equivalent
*For the Lysis Buffer, IC Diluent, and Blocking BUffer recommended for a specific DuoSet ELISA Development Kit, please see the product
The Human Total-FGF R2 alpha DuoSet IC ELISA is more sensitive than immunoprecipitation (IP)-Western blot analysis Lysates prepared from the human stomach cancer cell line (KatoIII) were diluted in series and analyzed by (A) IP-Western blot and (B) this DuoSet IC ELISA. IPs were performed using an anti-FGF R2 alpha monoclonal antibody and goat anti-mouse agarose. Immunoblots were incubated with a biotinylated anti-FGF R2 alpha monoclonal antibody. Bands were visualized with Streptavidin-HRP (Catalog # DY998) followed by chemiluminescent detection using WesternGlo Chemiluminescent Detection Substrate (Catalog # AR004). Human FGF R2 alpha can be detected by the Human Total-FGF R2 alpha DuoSet IC ELISA by using approximately 20 to 40 times less lysate than is needed for a conventional IP-Western blot.
Preparation and Storage
Background: FGFR2 alpha
FGF activity is mediated by a family of type I transmembrane tyrosine kinases, which undergo dimerization and autophosphorylation after ligand binding. Five distinct genes encode closely related FGF receptors, FGF R1 through 5. FGF Rs contain three Ig-like domains and a stretch of acidic residues between the first and second Ig-like domains. FGF R1, 2, 3, and -4 have a cytoplasmic split tyrosine-kinase domain, but FGF R5 does not. Multiple forms of FGF R1, 2, and 3 are generated by alternative splicing
Citations for Phospho-FGFR2 alpha DuoSet IC ELISA
R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.
Citations: Showing 1 - 2
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INCB054828 (pemigatinib), a potent and selective inhibitor of fibroblast growth factor receptors 1, 2, and 3, displays activity against genetically defined tumor models
Authors: PCC Liu, H Koblish, L Wu, K Bowman, S Diamond, D DiMatteo, Y Zhang, M Hansbury, M Rupar, X Wen, P Collier, P Feldman, R Klabe, KA Burke, M Soloviev, C Gardiner, X He, A Volgina, M Covington, B Ruggeri, R Wynn, TC Burn, P Scherle, S Yeleswaram, W Yao, R Huber, G Hollis
PLoS ONE, 2020;15(4):e0231877.
Sample Types: Cell Culture Media
Andrographolide enhanced 5-fluorouracil-induced antitumor effect in colorectal cancer via inhibition of c-MET pathway
Authors: M Su, B Qin, F Liu, Y Chen, R Zhang
Drug Des Devel Ther, 2017;11(0):3333-3341.
Sample Types: Cell Culture Supernates
Which phosphorylated sites are recognized by this assay?
This assay utilizes an anti-phosphorylated tyrosine monoclonal detection antibody, and it recognizes all phosphorylated tyrosine residues.
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