Recombinant bovine/porcine FGF2-G3 (154 aa) protein (Qk081)

Product: Qk081 Category:

£140.00£800.00

Qk081 bpFGF2-G3 vial image

Recombinant bovine/porcine FGF2-G3 (154 aa) protein is a thermostable engineered form of FGF-2. Qk081 is the 154 aa mature domain of FGF-2 (Qk056). FGF2-G3 has 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) cultivated meat and veterinary research applications including the expansion of bovine smooth muscles cells, bovine induced pluripotent, embryonic and mesenchymal stem cells.

Recombinant FGF2-G3 is used in B8 media (Kuo et al. 2020) 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.

Bovine/porcine thermostable FGF2-G3 (154 aa) has a molecular weight of 17.2 kDa. This protein is carrier-free and tag-free to ensure its purity with exceptional lot-to-lot consistency.

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£140.00£800.00

1000µg will be despatched as 2 x 500µg

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Summary:

  • High purity thermostable bovine/porcine FGF2-G3
  • 17.2 kDa (monomer)
  • >98%, by SDS-PAGE quantitative densitometry

  • 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 sterile-filtered water at >50 µg/ml, add carrier protein if desired, prepare single use aliquots and store frozen at -20 °C (short-term) or -80 °C (long-term).

Featured applications:

  • Bovine and porcine stem cell expansion and maintenance

  • Cellular agriculture and cultivated meat cell culture media optimization

Recombinant bovine/porcine FGF2-G3 activity was determined using the Promega serum response element luciferase reporter assay in transfected HEK293T cells. EC50 = 415 pg/ml (24 pM). Cells were treated in triplicate with a serial dilution of FGF2-G3 for 3 hours. Firefly luciferase activity was measured and normalized to the control Renilla luciferase activity. Data from Qk081 lot #204612.

Recombinant bovine/porcine FGF2-G3 154 aa migrates as a major band at approximately 17 kDa (monomer) in reduced (R) and non-reduced (NR) conditions. The dimeric form is also observed at approximately 34 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 Qk081 lot #204612.

Further quality assays

  • Mass spectrometry: single species with expected mass

  • Recovery from stock vial:  >95%

  • Endotoxin: <0.005 EU/μg protein (below level of detection)

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.

WT FGF-2 (Qk056) and FGF2-G3 (Qk081) were diluted in conditioned media and incubated at 37 °C. Samples were taken at 0 and 2 days. 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 Qk056 WT FGF-2 at 0 days (EC50 0.105 ng/ml) and 2 days (no EC50). Comparison between Qk081 FGF2-G3 at 0 hours (EC50 0.128 ng/ml) and 48 hours (EC50 0.103 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].

Additional resources

FAQ

What is FGF-2?

Fibroblast growth factor 2 (FGF-2), also known as basic fibroblast growth factor (bFGF) is a growth factor and signaling protein.

What does FGF-2 do?

FGF-2 is essential for normal embryonic development. It has roles in cell survival and proliferation, angiogenesis, tumorigenesis, wound healing and tissue repair.

What does FGF-2 bind to?

FGF-2 binds to and signals though all four of the FGF receptors FGFR1-4.

What is the function of the FGF receptor?

FGFRs phosphorylate specific tyrosine residues and activate the RAS-MAPK, PI3K-AKT, PLCγ, and STAT intracellular signaling pathways.

How is FGF-2 used in stem cell culture?

FGF-2 is used to maintain the pluripotency of stem cells in culture.

Our products are for research use only and not for diagnostic or therapeutic use. Products are not for resale.

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Qk081 bpFGF2-G3 vial imageRecombinant bovine/porcine FGF2-G3 (154 aa) protein (Qk081)
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