Which phosphorylated sites are recognized by this assay?

Name: Phospho-FGFR2 alpha DuoSet IC ELISA
Brand: R&D Systems
SKU: DYC684-2
Price: 470 USD

This assay utilizes an anti-phosphorylated tyrosine monoclonal detection antibody, and it recognizes all phosphorylated tyrosine residues.

Phospho-FGFR2 alpha DuoSet IC ELISA

Catalog #: DYC684-2
2 Citations Datasheet / COA / SDS

Catalog #	Availability	Size / Price	Qty
DYC684E
DYC684-5
DYC684-2

Ancillary Products Available

DY999B: Substrate Reagent Pack

DY999: Substrate Reagent Pack

The Human Total-FGF R2 alpha DuoSet IC ELISA is more sensitive than immunoprecipitation (IP)-Western blot analysis

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Product Details

Citations (2)

FAQs

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Summary Product Features Kit Content Other Reagents Required Data Examples Product Datasheets Preparation and Storage Background

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

Scientific Data

ELISA

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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

Product Datasheet

COA

SDS

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:

BBDS;BEK;BFR-1;CD332;CEK3;CFD1;ECT1;Fibroblast growth factor receptor;Fibroblast growth factor receptor 2;JWS;KGFR;K-SAM;Receptor protein-tyrosine kinase;TK14;TK25; 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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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, 2020-04-21;15(4):e0231877.
Species: Mouse
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, 2017-11-23;11(0):3333-3341.
Species: Human
Sample Types: Cell Culture Supernates

FAQs

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