Recombinant Human FGF-10 Heat Stable Protein, CF

Bioactivity of Recombinant Human FGF-10 Heat Stable Protein (Catalog # BT-FGF10HS) was measured in a cell proliferation assay using Ba/F3 mouse pro B cells transfected with human FGF RIIb. The ED50 for this effect is 4.00-60.0 ng/mL.

Recombinant Human FGF‑10 protein induces proliferation of Ba/F3 mouse pro-B cells transfected with human FGFR2b. (A) Recombinant Human FGF‑10 Heat Stable (Catalog # BT-FGF10HS) or (B) Recombinant Human FGF‑10 (345-FG) were either untreated or incubated at 37°C for 9 days in media. The heat-stable (HS) FGF‑10 retained similar bioactivity after incubation compared to the untreated HS protein, indicating that the HS protein has increased thermal stability. In contrast, the wild-type (WT) FGF‑10 protein showed a significant loss of activity following incubation, suggesting less thermal stability.

2 μg/lane of Recombinant Human FGF‑10 Heat Stable Protein (Catalog # BT-FGF10HS) was resolved with SDS-PAGE under reducing (R) and non-reducing (NR) conditions and visualized by Coomassie® Blue staining, showing bands at 19-23 kDa, under reducing conditions.

To evaluate the signal transduction response triggered by FGF-10, a HEK293 cell line that expresses a reporter for SEAP controlled by the Serum Response Element was utilized. Wild type FGF-10 (345-FG) and Heat Stable FGF-10HS (BT-FGF10HS) were incubated in DMEM media at 37°C for 48 or 72 hrs at concentrations of 600 ng/mL. Specific activity was assessed through a SEAP bioassay in HEK293 cells, wherein cells were treated with FGF-10 WT or FGF-10HS for 24 hours. The specific activity % of control was determined, with the control comprising FGF-10 stored at 4°C without any incubation.

Recombinant Human FGF R2 beta (IIIb) (Catalog # 665-FR) was captured on Biacore Sensor Chip CM5 via Recombinant Protein A/G/L (Catalog # NBP2-34985), and binding to Recombinant Human FGF 10 Heat Stable (Catalog # BT-FGF10HS) was measured at a concentration range between 0.0488 nM and 12.5 nM. The double-referenced sensorgram was fit to a two-state reaction binding model to determine the binding kinetics and affinity, with an affinity constant of KD=36.5 pM. (Biacore T200).

Recombinant Human FGF R2 beta (IIIb) (Catalog # 665-FR) was captured on Biacore Sensor Chip CM5 via Recombinant Protein A/G/L (Catalog # NBP2-34985), and binding to Recombinant Human FGF 10 (Catalog # 345-FG) was measured at a concentration range between 0.0488 nM and 12.5 nM. The double-referenced sensorgram was fit to a two-state reaction binding model to determine the binding kinetics and affinity, with an affinity constant of KD=36.2 pM. (Biacore T200).

Adult airway organoids were cultured in Cultrex UltiMatrix (Catalog # BME001-05) with either Recombinant Human FGF-10 Heat Stable Protein (HS, Catalog # BT-FGF10HS) or wild type Recombinant Human FGF-10 (WT, Catalog # 345-FG). FGF-10 Heat Stable demonstrates that it can support organoid growth as effectively as wild type in an extracellular matrix. Scale bar indicates 1 mm.

Adult airway organoids were cultured in Cultrex Synthetic Hydrogel (Catalog # CSH-RUO-01) with either Recombinant Human FGF-10 Heat Stable Protein (HS, Catalog # BT-FGF10HS) or wild type Recombinant Human FGF-10 Protein (WT, Catalog # 345-FG). FGF-10 Heat Stable Protein demonstrates that it can support organoid growth as effectively as wild type in a defined animal-free extracellular matrix. Scale bar indicates 1 mm.

• Reactivity: Hu
• Applications: Bioactivity
• Format: Carrier-Free

Recombinant Human FGF-10 Heat Stable Protein, CF Summary

• Details of Functionality: Measured in a cell proliferation assay using Ba/F3 mouse pro B cells transfected with human FGF RIIb. The ED₅₀ for this effect is 4.00-60.0 ng/mL.
• Source: E. coli-derived human FGF-10 protein
  Prorietary, engineered based on O15520.
• Protein/Peptide Type: Recombinant Proteins
• Purity: >90%, 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:
  • Bioactivity
• SDS-PAGE: 19-23 kDa, under reducing conditions.

Packaging, Storage & Formulations

• Storage: Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 3 months, -20 to -70 °C under sterile conditions after reconstitution.
• Buffer: Lyophilized from a 0.2 μm filtered solution in MOPS, Na₂SO₄, EDTA and DTT with Trehalose.
• Purity: >90%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining
• Reconstitution Instructions: Reconstitute at 250 μg/mL in water.

Notes

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

Alternate Names for Recombinant Human FGF-10 Heat Stable Protein, CF

• FGF10
• FGF-10
• fibroblast growth factor 10
• Keratinocyte growth factor 2
• KGF2
• KGF-2
• produced by fibroblasts of urinary bladder lamina propria

Background

Fibroblast Growth Factors including FGF-10 are known to be intrinsically unstable, and prone to unfold at 37°C, quickly rendering the growth factor inactive. Our de novo Heat Stable FGF-10 has been engineered for thermostability to maintain activity at 37℃ for at least 15 days. Heat-stable FGF-10 can enhance the consistency and efficacy of cell culture or organoid development. Heat Stable FGF-10 maintains sequence similarity, with >95% sequence homology to native FGF-10, making FGF-10 Heat Stable growth factor an easy replacement for culture.