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Recombinant FGF2-G3 (154 aa) protein (Qk053)
Recombinant FGF2-G3 (FGF2-STAB®) protein is a thermostable engineered form of FGF-2 (bFGF). Qk053 is the 154 aa mature domain of FGF-2 (Qk027) with nine amino acid substitutions to enhance stability without impacting bioactivity developed by Dvorak et al. 2018. This increases the functional half-life of the protein from <10 h (wild-type) to >7 days (FGF2-G3).
Recombinant FGF2-G3 is used in B8 media (Kuo et al. 2019) for weekend free, high homogeneity induced pluripotent stem cell culture. FGF2-G3 also has applications in chemically defined stem cell and organoid culture media, and cultured meat media development.
High purity 17 kDa bioactive FGF2-G3 protein, animal origin-free (AOF), carrier protein-free and tag free.
Orders are typically shipped same or next day (except Friday).
Easy world-wide ordering, direct or through our distributors.
1000µg will be despatched as 2 x 500µg
Fast and free shipping.
Buy online with secure credit card or purchase order.
For any questions, please email orders@qkine.com
Summary
High purity thermostable recombinant FGF2-G3 protein comprising 154 aa form of FGF-2 (Uniprot: P09038) with nine stabilizing amino acid substitutions
>98%, by SDS-PAGE quantitative densitometry
17 kDa
Expressed in E. coli.
Animal origin-free (AOF) and carrier protein-free.
Manufactured in our Cambridge, UK laboratories
Lyophilized from Tris/NaCl/CyS/mannitol
- Resuspend in water at >100 µg/ml, prepare single use aliquots, add carrier protein if desired, store frozen at -20 oC (short-term) or -80 oC (long-term)
Featured applications
- Expansion of induced pluripotent, embryonic and mesenchymal stem cells
- Cell expansion
Species neutral
Alternative proteins
Recombinant human FGF-2 (154 aa) protein (Qk027): wild-type form of this protein
Recombinant FGF-G3 (145 aa) protein (Qk052): 145 aa form of hyperstable FGF2-G3
Recombinant human FGF-2 (145 aa) protein (Qk025)
Recombinant zebrafish FGF-2 protein (Qk002)
This protein has recently been reformulated and will now arrive lyophilized. This change has been made to provide enhanced stability to ensure that we offer the best customer experience in terms of shipping and storage and to align with our company sustainability goals. Please visit our reconstitution page or contact customerservice@qkine.com for further information or to request the previous datasheet.
Bioactivity
Recombinant FGF2-G3 activity is determined using the Promega serum response element luciferase reporter assay (*) in transfected HEK293T cells. EC50 = 90 pg/ml (5.2 pM). Cells are treated in triplicate with a serial dilution of FGF2-G3 for 3 hours. Firefly luciferase activity is measured and normalized to the control Renilla luciferase activity. Data from Qk053 lot #104340. *Promega pGL4.33[luc2P/SRE/Hygro] #E1340
Purity
Recombinant FGF2-G3 migrates as a major band at 17 kDa in non-reducing (NR) conditions. The higher molecular weight minor band is the dimeric form. Upon reduction (R), only the 17 kDa band is visible. No contaminating protein bands are present. Purified recombinant protein (3 µg) was resolved using 15% w/v SDS-PAGE in reduced (+β-mercaptothanol, R) and non-reduced (NR) conditions and stained with Coomassie Brilliant Blue R250. Data from Qk053 batch #104340.
Further quality assays
Mass spectrometry: single species with expected mass
Analytical reversed-phase: single sharp peak
Endotoxin: <0.005 EU/μg protein (below level of detection)
Recovery from stock vial >95%
We are a company founded and run by scientists to support innovation in stem cell biology and regenerative medicine. To enhance reliability and reproducibility in your applications, all our products are exceptionally high purity, with complete characterisation and bioactivity analysis on every lot.
Product manufactured under license: International Patent Application No. PCT/EP2016/073567 and U.S. Patent No. 11,746,135
Wild-type FGF-2 (Qk027) and FGF2-G3 (Qk053) have equivalent bioactivity
FGF2-G3 activity is determined using the Promega serum response element luciferase reporter assay (*) in transfected HEK293T cells. Cells are treated in triplicate with a serial dilution of FGF2-G3 for 3 hours. Firefly luciferase activity is measured and normalized to the control Renilla luciferase activity. *Promega pGL4.33[luc2P/SRE/Hygro] #E1340
‘After a long day running samples and analysing data, it was very exciting to see just how stable the FGF2-G3 proteins were. They clearly have more endurance than me!’
– Kerry, Qkine Senior Scientist
Hyperstable FGF2-G3 retains activity after pre-incubation with conditioned media at 37°C
+24 hours
+48 hours
+ 72 hours
FGF2-G3 (Qk053) WT FGF2 (Qk027)
WT FGF-2 (Qk027) and FGF2-G3 (Qk053) were diluted in conditioned media and incubated at 37oC. Samples were taken at 24 h intervals and FGF-2 activity was assayed in triplicate using the Promega serum response element luciferase reporter assay (*) in transfected HEK293T cells. Firefly luciferase activity was normalized to the control Renilla luciferase activity (F/R).
Recombinant FGF2-G3 protein background
FGF-2 (also known as basic FGF or bFGF) is an essential growth factor for maintaining human embryonic stem cell (hESC) and induced pluripotency stem cell (iPSC) pluripotency in feeder-free and chemically defined stem cell media. It is a core component of widely adopted media including mTESR (Ludwig et al. 2006), StemPRO (Wang et al. 2007) and E8 (Chen et al. 2011). However, FGF-2 is inherently unstable and prone to proteolytic degradation and aggregation. This fundamental biochemical instability, and therefore low half-life in culture media (<10 h), is an important contribution to the need for frequent media changes and challenges in improving homogeneity during stem cell proliferation and subsequent differentiation.
To improve the stability of FGF-2 for stem cell media and regenerative medicine applications, Dvorak and colleagues at Masaryk University used computer-assisted protein engineering to identify an optimal set of nine amino acid substitutions that stabilize FGF-2. These substitutions were designed to avoid structural changes to the FGF receptor 1 (FGFR1) and FGF receptor 2 (FGFR2) binding interface. This thermostable FGF-2 is known as FGF2-G3, or FGF2-STAB® (Dvorak et al. 2017). The biological activity of wild-type FGF-2 is <50% after 10h incubation with conditioned media. In contrast, no reduction in FGF2-G3 biological activity is observed after >7 days incubation with conditioned media at 37oC. Both FGF2-G3 and wild-type FGF-2 maintain hESC pluripotency and expression of pluripotency markers Oct-4 and nanog with equivalent efficacy (Dvorak et al. 2017).
In 2020, Paul Burridge and colleagues at Northwestern University, Chicago, published a protocol for B8 media. This iPSC maintenance media uses thermostable FGF2-G3, along with optimization of media component concentration and composition to reduce media cost and facilitate weekend-free stem cell culture regimes (Kuo et al. 2020 and updated in Lyra-Leite et al. 2020).
To manufacture and provide recombinant FGF2-G3 for stem cell culture, including use in B8 media and emerging applications such as cellular agriculture, Qkine has licensed the patented FGF2-G3 technology from Enantis/Masaryk University. We have combined the excellent science behind the FGF2-G3 technology with our protein manufacture expertise to provide gold standard protein for use in cell culture media. We have removed His tags present in the academic forms of the protein for FGF2-G3 (Qk053), as these may cause issues for scientists looking to translate discoveries to clinical or scale-up applications, and introduce unnecessary scientific uncertainties.
The 9 aa pro-segment of FGF2 present in the 154 aa FGF-2 is not required for biological activity and is thought to have roles in the localization of FGF-2 in vivo.
We wanted to allow scientists to compare directly with their existing FGF-2, so we introduced the nine amino acid substitutions into the 145 aa form of FGF-2 to make Qk052 FGF2-G3 (145 aa). The EC50 of the two forms of FGF2-G3 are 90 pg/ml and 47 pg/ml for the 154 aa and 145 aa forms respectively. As observed previously, the half-life is extended to >3 days.
Key papers
Additional resources
Publications using recombinant FGF2-G3 (154 aa) protein (Qk053)
Modeling the selective growth advantage of genetically variant human pluripotent stem cells to identify opportunities for manufacturing process control.
In Cytotherapy in April 2024 by Beltran-Rendon, C., Price, C. J. et al.
Ephrin-A2 and Phosphoantigen-Mediated Selective Killing of Medulloblastoma by γδ T Cells Preserves Neuronal and Stem Cell Integrity
bioRxiv preprint on 13 October 2024 by Boutin, L. et al.
mTOR activity paces human blastocyst stage developmental progression
In Cell on 26 September 2024 by Lyer, D. P., Khoei, H. H. et al.
A chemically defined and xeno-free hydrogel system for regenerative medicine.
Preprint on 2 June 2024 by Ong, J. Gibbons, G., Siang, L. Y. et al.
Feeder-free culture of human pluripotent stem cells drives MDM4-mediated gain of chromosome 1q.
In Stem Cell Reports on 13 August 2024 by Stavish, D. et al.
Cytogenetic resource enables mechanistic resolution of changing trends in human pluripotent stem cell aberrations linked to feeder-free culture.
Preprint on 21 September 2023 by Stavish, D. et al.
Refined home-brew media for cost-effective, weekend-free hiPSC culture and genetic engineering.
In Open Research Europe 2024 by Truszkowski, L., Bottini, S., Bianchi, S. et al.
Development of an orthotopic medulloblastoma zebrafish model for rapid drug testing.
Preprint on 121 February 2024 by van Bree, N., Oppelt, A. et al.
Our products are for research use only and not for diagnostic or therapeutic use. Products are not for resale.
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