Recombinant Human FGF-10 Heat Stable Protein, CF Summary
- R&D Systems E. coli-derived Recombinant Human FGF-10 Heat Stable Protein (BT-FGF10HS)
- Quality control testing to verify active proteins with lot specific assays by in-house scientists
- All R&D Systems proteins are covered with a 100% guarantee
Product Specifications
Prorietary, engineered based on O15520.
Product Datasheets
Carrier Free
CF stands for Carrier Free (CF). We typically add Bovine Serum Albumin (BSA) as a carrier protein to our recombinant proteins. Adding a carrier protein enhances protein stability, increases shelf-life, and allows the recombinant protein to be stored at a more dilute concentration. The carrier free version does not contain BSA.
In general, we advise purchasing the recombinant protein with BSA for use in cell or tissue culture, or as an ELISA standard. In contrast, the carrier free protein is recommended for applications, in which the presence of BSA could interfere.
BT-FGF10HS
| Formulation | Lyophilized from a 0.2 μm filtered solution in MOPS, Na2SO4, EDTA and DTT with Trehalose. |
| Reconstitution | Reconstitute at 250 μg/mL in water. |
| Shipping | The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. |
| Stability & Storage: | Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
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Scientific Data
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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.
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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.
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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.
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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.
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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).
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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).
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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.
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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.
Reconstitution Calculator
Background: FGF-10
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.
FAQs
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