Recombinant Human FGFR2 kinase domain His-tag Protein, CF

Recombinant Human FGFR2 kinase domain His-tag (Catalog # 11812-FR) is measured by its ability to transfer phosphate from adenosine triphosphate (ATP) to a peptide substrate.

• Applications: Enzyme Activity
• Format: Carrier-Free

Recombinant Human FGFR2 kinase domain His-tag Protein, CF Summary

• Details of Functionality: Measured by its ability to transfer phosphate from adenosine triphosphate (ATP) to a peptide substrate.
  The specific activity is >75 pmol/min/μg, as measured under the described conditions.
• Source: Human embryonic kidney cell, HEK293-derived human FGFR2 protein
  Met459-Leu772, with a N-terminal 6-His tag
• N-terminal Sequence: Protein identity confirmed by mass spectrometry
• Protein/Peptide Type: Recombinant Proteins
• Purity: >80%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining
• Endotoxin Note: <0.10 EU per 1 μg of the protein by the LAL method.

Applications/Dilutions

• Dilutions:
  • Enzyme Activity
• Theoretical MW: 37 kDa.
  Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.
• SDS-PAGE: 35-39 kDa, under reducing conditions.

Packaging, Storage & Formulations

• Storage: Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 6 months from date of receipt, -20 to -70 °C as supplied.
  • 3 months, -20 to -70 °C under sterile conditions after opening.
• Buffer: Supplied as a 0.2 μm filtered solution in Tris, NaCl, DTT and Glycerol.
• Purity: >80%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining
• Assay Procedure:
  • Assay Buffer: 50 mM Tris, 20 mM MgCl₂, 5 mM MnCl₂, 0.1 mg/mL BSA, pH 7.5
  • Recombinant Human FGFR2 kinase domain His-tag (rhFGFR2) (Catalog # 11812-FR)
  • Poly (4:1 Glu, Tyr) Peptide, 1 mg/mL stock in 25 mM Tris, pH 7.5
  • Adenosine Triphosphate (ATP), 10 mM stock in deionized water
  • ADP-Glo^TM Kinase Assay Kit (Promega)
  • White 96-well Plate
  • Plate Reader with Luminescence Read Capability
  1. Dilute rhFGFR2 to 10 µg/mL in Assay Buffer.
  2. Prepare Substrate Mixture containing 200 µM ATP and 0.6 mg/mL Poly (4:1 Glu, Tyr) Peptide in Assay Buffer.
  3. Combine equal volumes of 10 µg/mL rhFGFR2 and Substrate Mixture. Create a Substrate Control containing equal volumes of Assay Buffer and Substrate Mixture.
  4. Incubate at room temperature for 40 minutes in the dark.
  5. After incubation, transfer 10 µL of each reaction to a plate.
  6. Terminate the reaction and deplete the remaining ATP by adding 10 µL of ADP-Glo^TM Reagent (supplied in kit) to all wells.
  7. Incubate at room temperature for 40 minutes in the dark.
  8. Add 20 µL Kinase Detection Reagent (supplied in kit) to all wells.
  9. Incubate at room temperature for 30 minutes in the dark.
  10. Read plate in Luminescence endpoint mode.
  11. Calculate specific activity:

  Specific Activity (pmol/min/µg) = (Adjusted Luminescence* (RLU) x Conversion Factor** (pmol/RLU)) / (Incubation time (min) x amount of enzyme (µg))

  *Adjusted for Substrate Control
  **Derived from ADP-Glo^TM Kinase Assay Kit protocol (Promega)
  Per Reaction:
  • rhFGFR2: 5 μg/mL
  • ATP: 100 µM
  • Poly (4:1 Glu, Tyr) Peptide: 0.3 mg/mL

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for Recombinant Human FGFR2 kinase domain His-tag Protein, CF

• BBDS
• BEK
• BFR-1
• CD332 antigen
• CD332
• CEK3
• CFD1
• craniofacial dysostosis 1
• EC 2.7.10
• EC 2.7.10.1
• ECT1
• FGF R2
• FGFR2
• fibroblast growth factor receptor 2
• FLJ98662
• Jackson-Weiss syndrome
• JWS
• Keratinocyte growth factor receptorreceptor like 14
• KGFR
• KSAM
• K-sam
• soluble FGFR4 variant 4
• TK14
• TK25

Background

Fibroblast growth factor receptor 2 (FGFR2), also known as Keratinocyte growth factor receptor (KGFR), is one of four highly conserved tyrosine protein kinase FGFR receptors within the tyrosine protein kinase family that differ in ligand affinity and tissue distribution. FGFR2, like other receptor members in this family, is comprised of an extracellular domain (ECD) with immunoglobulin-like segments that binds its target cytokine FGF and heparin sulfate to induce dimerization and autophosphorylation, a single-pass transmembrane domain, and a cytosolic kinase domain that triggers downstream signaling cascades through multiple pathways (1, 2). Like the structure of other tyrosine kinases, the kinase domain contains the conserved catalytic domain with an ATP binding site within the cleft of two lobes at the N-terminal and C-terminal regions as well as an activation loop at the surface between the two lobes (3). FGFR2 has two naturally occurring isoforms, FGFR2b and FGFR2c, that are created by splicing of the third immunoglobulin-like domain and lead to tissue specific expression within epithelial and mesenchyme derived tissues respectively to bind different FGFs (4, 5). Missense mutations of FGFR2 that cause gain of function cause several congenital skeletal disorders including Apert, Crouzon, and Pfeiffer syndromes (6, 7). In addition, FGFR2 regulates signaling pathways that promote cell proliferation, inhibition of apoptosis, and angiogenesis (5). FGFR2 has been reported to be overexpressed or mutated in many tumors including brain, breast, cholangiocarcinoma, gastric, lung, ovarian and uterine, some of which have been further correlated with poor prognosis (5, 8-15). Inhibition of FGFR2 has been identified as a promising cancer therapeutic strategy and there is significant ongoing research in the development of drugs that inhibit FGFR2 using multikinase and more selective ATP-binding site small molecule and antibody drugs (2, 5, 14, 16).

1. Hubbard, S.R. and J.H. Till (2000) Annu. Rev. Biochem. 69:373.
2. Katoh, M. et al. (2024) Nature Rev. Clin. Oncol. 21:312. 
3. Dai, S. et al. (2019) Cell. 8:614.
4. Orr-Urtreger A. et al. (1993) Developmental Biology. 158:475.
5. Lages Dos Santos,J. et al. (2025) Medicina 61:1890. 
6. Pollock, P.M. et al. (2007) Oncogene 26:7158.
7. Tuzon, C.T. et al. (2019) Curr. Osteoporos Rep. 17:138.
8. Shoji H. et al. (2015) Anticancer Res. 35:5055.
9. Liu, G. et al. (2017) Tumour Biol. 39:1010428317707424.
10. Mahipal, A. et al. (2019) Cancer Treat. Rev. 78:1.
11. Georgescu, M.M. et al. (2021) Acta Neuropathol. Commun. 9:69.
12. Jogo, T. et al. (2021) Clin. Cancer Res. 27:5619.
13. Pacini, L. et al. (2021) Cells 10:1154.
14. Francavilla, C. and C.S. O'Brien (2022) Open Biol. 12:210373.
15. Vogel, A. et al. (2023) Annu. Rev. Med. 74:293.
16. Zingg, D. et al. (2022) Nature 608:609.