The total FGFR2 kit monitors the cellular FGFR2 expression level and can be used as a normalization assay for the phospho-FGFR kit.
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  • Ease-of-use Ease-of-use
  • Highly specific Highly specific
The total FGFR2 kit monitors the cellular FGFR2 expression level and can be used as a normalization assay for the phospho-FGFR kit.
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Overview

The Total FGFR2 cellular assay monitors total FGFR and is used as a normalization assay with the phospho-FGFR kit. Mutations in FGFR2 have been associated with cancer (breast, melanoma) and abnormal bone development (cranosynostosis syndromes).

Benefits

  • VALIDATED ON KATO III CELLS
  • SPECIFICITY
  • DATA NORMALIZATION

Total-FGFR2 assay principle

The Total-FGFR2 assay quantifies the expression level of FGFR2 in a cell lysate. Contrary to Western Blot, the assay is entirely plate-based and does not require gels, electrophoresis or transfer. The Total-FGFR2 assay uses two labeled antibodies: one coupled to a donor fluorophore, the other to an acceptor. Both antibodies are highly specific for a distinct epitope on the protein. In presence of FGFR2 in a cell extract, the addition of these conjugates brings the donor fluorophore into close proximity with the acceptor and thereby generates a FRET signal. Its intensity is directly proportional to the concentration of the protein present in the sample, and provides a means of assessing the protein’s expression under a no-wash assay format.
Total FGFR2 assay principle

Total-FGFR2 2-plate assay protocol

The 2 plate protocol involves culturing cells in a 96-well plate before lysis then transferring lysates to a 384-well low volume detection plate before adding Total FGFR2 HTRF detection reagents. This protocol enables the cells' viability and confluence to be monitored.
Total FGFR2 2-plate assay protocol

Total-FGFR2 1-plate assay protocol

Detection of total FGFR2 with HTRF reagents can be performed in a single plate used for culturing, stimulation and lysis. No washing steps are required. This HTS designed protocol enables miniaturization while maintaining robust HTRF quality.
Total FGFR2 1-plate assay protocol

Detection phospho and total FGFR2 on Kato-III cells

Kato-III cells were plated in 96-well plate and incubated overnight. Cells were treated with increasing concentration of FIIN-1 hydrochloride compound for 6h at 37 °C - 5% CO2. After stimulation, cells were washed and lysed with lysis buffer#2 1X for 30 min at RT under gentle shaking. 16 µL of lysate were transferred into a 384-well sv white microplate and 4 µL of the HTRF phospho FGFR2 or total FGFR2 detection reagents were added. The HTRF signal was recorded after a overnight incubation at room temperature.
Detection phospho and total FGFR2 on Kato-III cells
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Cisbio lysis buffer compatibility

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Species compatibility

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Product Insert FGFR2 total Kit / 63ADK057PEG-63ADK057PEH

63ADK057PEG-63ADK057PEH - Product Insert

Cisbio Product Catalog 2019

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Get the brochure about technology comparison. - Brochures

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