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Recombinant FGF-2 TOP® (Thermostability Optimized)

Animal component-free, Endotoxin-free, Biorisk-free / Next-generation Growth Factors for Stem Cells and Cell-based applications

Recombinat FGF-2 TOP® (formerly STAB) is a stabilized growth factor that offers a novel way to grow FGF-2 dependent cell cultures more efficiently, with fewer media changes. FGF-2 TOP® retains full biological activity even after twenty days. The stable level of FGF-2 in culture allows for a more homogenous, undifferentiated stem cell culture, while saving researchers valuable time and money because repeated supplementation with FGF-2 and a daily medium change is not required.

FGF-2 TOP® is the 155 aa mature domain of FGF-2, tag-free 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 (FGF-2 TOP®) in cell culture conditions at 37ºC.

Product Overview

Recombinant FGF-2 TOP® is produced by recombinant expression of the human sequence of FGF-2 in plant seeds of Camelina sativa. Modifications to the sequence for improved expression, bioactivity and stability are applied but are proprietary. None of the components or raw materials employed for the production of FGF-2 TOP® are derived or extract from animal or human origin.

 

Uniquely Engineered for Stem Cell Production:

FGF-2 TOP® engineered with a novel nine amino acid substitution to improve stability is thermostabilized growth factor that enables more efficient FGF-2-dependent cell cultures with fewer media changes.

 
Wild Type FGF-2 FGF-2 TOP®

 

Product Information:

   
Alternative Names
FGF-2 G3, bFGF STAB, Thermostable FGF-2
Accession Number
Amino Acid Sequence
AAGSITTLPALPEDGGSGAF PPGHFKDPKRLYCKNGGFFLRIHPDGRVDG
VREKSDPHIKLQLQAEERGVVSIKGVCANRYLAMKEDGRLLASKCVTDEC
FFFERLESNNYNTYRSRKYTSWYVALKRTGQYKLGSKTGPGQKAILFLPMSAKS
* Core Biogenesis FGF-2 TOP® is the 154 aa mature domain of FGF-2 with nine amino acid substitutions
Molecular mass
around 19.0kDa
Origin
Plant seeds of Camelina Sativa
Species
Engineered sequence
Similarity
Human (99%), Bovine (99%), Porcine (99%), Mouse (94%)

 

Specifications: 

Criteria Result
Purity
≥ 95% measured by SDS page
Bioactivity
The specific activity is EC50 0.5-1.0 ng/ml. Determined by the ability to promote the proliferation of NIH/3T3 cells cultured in adherent condition.
Formulation
Lyophilized powder
Endotoxin levels
≤ 0.005 ng/ug (≤ 0.005EU/ug of protein) as determined by Limulus Amebocyte Lysate Assay.
Animal / Human components
Free

Superior Bioactivity

 

 

FGF-2 TOP® (Thermostability Optimized) has an expected ED50 of less than 0.1 ng/mL, as determined by NIH-3T3 cell proliferation assay. The ED50 value of FGF-2 TOP® is about 5-times lower than that of the wild-type FGF-2, as determined by a NIH/3T3 proliferation assay. Under the same experimental conditions, less FGF-2 TOP® is needed to achieve a given effect on cell proliferation, thanks to the higher stability and prolonged half-life of FGF-2 TOP®. Thanks to this competitive advantage, lower dosage of FGF-2 are required in your cell culture applications (up to 10x).

Superior Thermostability

 

FGF-2 TOP® is a stabilized growth factor that offers a novel way to grow FGF-2 dependent cell cultures more efficiently, with fewer media changes. FGF-2 TOP® retains full biological activity even after twenty days at 37°C. The stable level of FGF-2 in culture allows for a more homogenous, undifferentiated pluripotent stem cell cultures, and consistent downstream differentiations, while saving researchers valuable time and money because repeated supplementation with FGF-2 and a daily medium change is not required.

 

 

Reduced Costs and “Weekend-Free” Feeding

Realize tangible benefits of FGF-2 TOP® protein stability: significant reductions in the amount of media required to feed cells, as well as reductions in the number of feedings, saving labor costs and obviating the need for inconvenient weekend feedings. Refer to the example feeding schedule and cost comparison of wild-type versus TOP®:

 

 

 

Publications

• FGF2-G3 was developed by Dvorak et al., 2018. Biotechnology and Bioengineering

Dvorak, P., Bednar, D., Vanacek, P., Balek, L., Eiselleova, L., Stepankova, V., Sebestova, E., Kunova Bosakova, M., Konecna, Z., Mazurenko, S. and Kunka, A., 2018. Computer‐assisted engineering of hyperstable fibroblast growth factor 2. Biotechnology and bioengineering115(4), pp.850-862.

 

• B8 media protocol established by Kuo et al., 2020. Stem Cell Reports 

Kuo, H.H., Gao, X., DeKeyser, J.M., Fetterman, K.A., Pinheiro, E.A., Weddle, C.J., Fonoudi, H., Orman, M.V., Romero-Tejeda, M., Jouni, M. and Blancard, M., 2020. Negligible-cost and weekend-free chemically defined human iPSC culture. Stem Cell Reports14(2), pp.256-270.

 

• updated in Lyra-Leite et al., 2021. STAR Protocols 

Lyra-Leite, D.M., Fonoudi, H., Gharib, M. and Burridge, P.W., 2021. An updated protocol for the cost-effective and weekend-free culture of human induced pluripotent stem cells. STAR protocols2(1), p.100213.

 

• Review of FGF 2 stabilization by Benington et al., 2020. Pharmaceutics

Benington, L., Rajan, G., Locher, C. and Lim, L.Y., 2020. Fibroblast growth factor 2—A review of stabilisation approaches for clinical applications. Pharmaceutics12(6), p.508.

 

• Simple and effective serum-free medium for sustained expansion of bovine satellite cells for cell cultured meat, Kaplan et al., 2021.

Stout, A.J., Mirliani, A.B., Rittenberg, M.L., Shub, M., White, E.C., Yuen Jr, J.S. and Kaplan, D.L., 2022. Simple and effective serum-free medium for sustained expansion of bovine satellite cells for cell cultured meat. Communications Biology5(1), p.466.

Download

 Thermostable Recombinant FGF-2 (FGF-2 STAB) (Product Information)

 Thermostable Recombinant FGF-2 (FGF-2 STAB) (Brochure)

 

Related Product: 

 FGF-2 Human (Product Information)

 FGF-2 Bovine (Product Information)

Ordering Information

Product Storage Cat.No. PKG Size Price  
FGF-2 TOP® -20°C PL9-50 50 µg 366.00
Buy
FGF-2 TOP® -20°C PL9-1000 1 mg 886.00
Buy
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