Technote | FGF-2 (Qk502Qk027)
Thermostable FGF2-G3 protein remains stable and highly bioactive in culture media for 7 days
Fibroblast growth factor 2 (FGF-2) is an essential growth factor for regulating several biological processes and a critical component for stem cell maintenance in feeder-free media. Qkine FGF2-G3 (Qk052) was engineered with nine amino acid substitutions to stabilize its structure and expressed in E. coli creating the only animal-free, protein-tag-free stabilized FGF2-G3 available commercially. The enhanced stability makes FGF2-G3 a highly bioactive and thermostable (heat stable) engineered form of FGF-2, resulting in improved culture conditions with less frequent media changes.
FGF2-G3 stability and bioactivity
Thermostable FGF2-G3 145 aa maintains bioactivity for >7 days under cell culture conditions. Bioactivity was determined using the Promega serum response element luciferase reporter assay in transfected HEK293T cells. FGF2-G3 145 aa or WT FGF-2 154 aa (Qk027) were incubated in media at 37°C for 0, 2 or 7 days then cells were treated in triplicate with a serial dilution for 3 hours. Firefly luciferase activity was measured and normalized to the control Renilla luciferase activity.
FGF2-G3 provides a reliable source of animal-free, highly bioactive, and thermostable FGF-2 for stem cell culture and FGF2-dependent applications. The use of stable FGF2-G3 results in enhanced stem cell cultures with less frequent media changes improving self-renewal maintenance and enhancing the reproducibility of subsequent differentiation the reproducibility of subsequent differentiation regenerative medicine applications.
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