Phospho-FGFR2 alpha DuoSet IC ELISA

Catalog # Availability Size / Price Qty
DYC684E
DYC684-5
DYC684-2
Ancillary Products Available
The Human Total-FGF R2 alpha  DuoSet IC ELISA is more sensitive than immunoprecipitation (IP)-Western blot analysis
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Citations (2)
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Phospho-FGFR2 alpha DuoSet IC ELISA Summary

Assay Type
Solid Phase Sandwich ELISA
Format
96-well strip plate
Sample Volume Required
Cell lysates (100 µL)
Sufficient Materials
Kits available for two, five, or fifteen 96-well plates*
Specificity
Please see the product datasheet

* Provided that the recommended microplates, buffers, diluents, substrates and solutions are used, and the assay is run as summarized in the Assay Procedure provided.

This DuoSet IC ELISA contains the basic components required for the development of sandwich ELISAs to measure tyrosine-phosphorylated human FGF R2 alpha in cell lysates. An immobilized capture antibody specific for FGF R2 alpha binds both phosphorylated and unphosphorylated FGF R2 alpha. After washing away unbound material, a biotinylated detection antibody is used to detect only phosphorylated receptor, utilizing a standard HRP format.

Product Features

  • 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

Kit Content

  • 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

Lysis Buffer*

IC Diluent*

Blocking Buffer*


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

Data Example

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.

Product Datasheets

Preparation and Storage

Shipping
The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage
Store the unopened product at 2 - 8 °C. Do not use past expiration date.

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

Long Name:
Fibroblast Growth Factor Receptor 2 alpha
Entrez Gene IDs:
2263 (Human); 14183 (Mouse)
Alternate Names:
FGF R2a; FGFR2 alpha

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.

2 Citations: Showing 1 - 2
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  1. 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.
    Species: Mouse
    Sample Types: Cell Culture Media
  2. 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.
    Species: Human
    Sample Types: Cell Culture Supernates

FAQs

  1. 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.

View all ELISA FAQs

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