Recombinant bovine/porcine FGF2-G3 (145 aa) protein (Qk080-FG)

Name: Recombinant bovine/porcine FGF2-G3 (145 aa) protein (Qk080-FG)
SKU: Qk080-FG
Availability: InStock

Recombinant bovine/porcine FGF2-G3 protein is a thermostable engineered form of FGF-2 (bFGF) with an increased functional half-life from <10 h (wild-type) to >7 days (FGF2-G3). Bovine FGF2-G3 is used for the development of optimized serum-free culture media for species-specific bovine (cow) and porcine (pork) cultivated meat and veterinary research applications, including the expansion of bovine and porcine smooth muscle cells, bovine and porcine induced pluripotent, embryonic and mesenchymal stem cells.

Recombinant FGF2-G3 is used in B8 media (Kuo et al. 2019) and Beefy-9 media (Stout et al. 2023) for weekend free, high homogeneity induced pluripotent stem cell culture. FGF2-G3 also has applications in chemically defined stem cell and organoid culture media.

Qkine high quality food grade bovine/porcine FGF2-G3 has a molecular weight of 16.3 kDa. This protein is animal origin-free, carrier-free and tag-free to ensure purity with exceptional lot-to-lot consistency.

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1mg will be despatched as 2 x 500µg

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Description

Summary

High purity thermostable bovine/porcine FGF2-G3
>98%, by SDS-PAGE quantitative densitometry
Source: Expressed in E. coli
16.3 kDa (Monomer)
Animal origin-free (AOF) and carrier protein-free
Manufactured in Cambridge, UK
Lyophilized from Tris/NaCl/CyS/mannitol
Resuspend in water at >100 µg/mL, prepare single-use aliquots, add carrier protein if desired, and store frozen at -20^oC or -80^oC

Handling and storage FAQ

Featured applications

High quality food grade recombinant protein
Expansion of bovine and porcine induced pluripotent, embryonic and mesenchymal stem cells
Cell expansion
Proliferation of fibroblasts and endothelial cells
Tissue repair and regeneration

Alternative protein names

Basic fibroblast growth factor
bFGF
FGF-β
FGF2
Fibroblast growth factor-basic
HBGF-2
FGF2-G3
FGF2-STAB
FGF2-STAB3

Species reactivity

Bovine
Porcine

Related discovery kit

FGF-2 species-specific discovery kit (Qk501)

Bioactivity

Recombinant bovine/porcine FGF2-G3 activity is determined using the Promega serum response element luciferase reporter assay (*) in transfected HEK293T cells. EC50 = 217 pg/ml (13 pM). Cells are treated in triplicate with a serial dilution of bovine/porcine FGF2-G3 for 3 hours. Firefly luciferase activity is measured and normalized to the control Renilla luciferase activity. Data from Qk080 lot #204611. *Promega pGL4.33[luc2P/SRE/Hygro] #E1340

Purity

Recombinant bovine/porcine FGF2-G3 145aa migrates as a major band at approximately 16 kDa (monomer) in reduced (R) and non-reduced (NR) conditions. The dimeric form is also observed at approximately 32 kDa in the non-reduced condition. No contaminating protein bands are present. The purified recombinant protein (7 µg) was resolved using 15% w/v SDS-PAGE in reduced (+β-mercaptoethanol, R) and non-reduced (NR) conditions and stained with Coomassie Brilliant Blue R250. Data from Qk080 lot #204611.

Further quality assays

Mass spectrometry, single species with the expected mass
Endotoxin: <0.005 EU/μg protein (below the level of detection)
Recovery from stock vial: >95%

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We are a company founded and run by scientists to provide a service and support innovation in stem cell biology and regenerative medicine. All our products are exceptionally high purity, with complete characterisation and bioactivity analysis on every lot.

Wild-type bovine/porcine FGF-2 and bovine/porcine FGF2-G3 have equivalent bioactivity

FGF2-G3 WT FGF-2

Bovine/porcine 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. Qk080 bovine/porcine FGF2-G3 (EC50 0.190 ng/ml) and Qk040 WT FGF-2 (EC50 0.237 ng/ml). *Promega pGL4.33[luc2P/SRE/Hygro] #E1340

Compared to wild-type bovine/porcine FGF-2, thermostable bovine/porcine FGF2-G3 retains activity after pre-incubation with conditioned media at 37°C for 48 hours

Wild-type FGF-2

FGF2-G3

0 hours + 48 hours

WT FGF-2 (Qk040) and FGF2-G3 (Qk080) were diluted in conditioned media and incubated at 37 °C. Samples were taken at 0 and 48 hours. FGF-2 activity was assayed in triplicate using the Promega serum response element luciferase reporter assay (*) in transfected HEK293T cells. Results were then normalized to the maximum response for 0 hours. Comparison between Qk040 WT FGF-2 at 0 hours (EC50 0.237 ng/ml) and 48 hours (EC50 1.75 ng/ml. Comparison between Qk080 FGF2-G3 at 0 hours (EC50 0.464 ng/ml) and 48 hours (EC50 0.606 ng/ml).

Protein background

FGF-2 (also known as basic FGF or bFGF) is an essential growth factor for maintaining embryonic stem cell (ESC) 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 [1], StemPRO [2] and E8 [3]. 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.

Bovine/porcine FGF2-G3 can be a core ingredient in Beefy-9 media, established by Stout et al. in 2023. The innovation of adding recombinant albumin to B8 media has yielded a groundbreaking advancement in cell culture for the growth of bovine satellite cells, resulting in the development of Beefy-9 media. This novel medium is the first of its kind, meeting fundamental criteria to serve as a foundational medium for cultured meat production. One key feature includes the complete absence of animal-derived components, ensuring animal-component-free culture conditions. This departure from traditional serum-containing media, which often rely on animal-derived additives, distinguishes Beefy-9’s commitment to ethical and sustainable practices in cell culture [4-6].

Background references

[1] Ludwig, Tenneille E., et al. ‘Feeder-Independent Culture of Human Embryonic Stem Cells’. Nature Methods, vol. 3, no. 8, Aug. 2006, pp. 637–46. doi: 10.1038/nmeth902

[2] Wang, Linlin, et al. ‘Self-Renewal of Human Embryonic Stem Cells Requires Insulin-like Growth Factor-1 Receptor and ERBB2 Receptor Signaling’. Blood, vol. 110, no. 12, Dec. 2007, pp. 4111–19. doi: 10.1182/blood-2007-03-082586

[3] Beers, Jeanette, et al. ‘Passaging and Colony Expansion of Human Pluripotent Stem Cells by Enzyme-Free Dissociation in Chemically Defined Culture Conditions’. Nature Protocols, vol. 7, no. 11, 2012, pp. 2029–40. doi: 10.1038/nprot.2012.130

[4] Stout, A. J. et al. A Beefy-R culture medium: Replacing albumin with rapeseed protein isolates. Biomaterials 296, 122092 (2023). https://doi.org/10.1016/j.biomaterials.2023.122092

[5] Kuo, H. H. et al. Negligible-Cost and Weekend-Free Chemically Defined Human iPSC Culture. Stem Cell Reports 14, 256–270 (2020). doi: 10.1016/j.stemcr.2019.12.007

[6] Mcaleer, C. W., Rumsey, J. W., Stancescu, M. & Hickman, J. J. Functional myotube formation from adult rat satellite cells in a defined serum-free system. Biotechnol. Prog. 31, 997–1003 (2015). doi: 10.1002/btpr.2063

Qkine high quality food grade products are intended solely for use as food processing aids, ex vivo cell manufacturing and research use. Not for direct human consumption, therapeutic or diagnostic use.

The authorization of novel foods, including the use of growth factors as food processing aids, is regulated by regional government agencies. The use of our products as food processing aids in novel foods requires the end-user to obtain the necessary regulatory approvals.

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