Recombinant human TGF-β3 protein

Summary

• High purity human TGF-β3 (Uniprot: P10600)
• 12.7 kDa (monomer) 25.4 kDa (dimer)
• >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 acetonitrile, TFA
• Resuspend in 10 mM HCl (Reconstitution solution A) 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

• Stem cell expansion and maintenance
• Induced pluripotent and embryonic stem cell differentiation and maintenance
• iPSC-derived mesoderm differentiation

TGF-β3 activity is determined using a TGF-β3 responsive firefly luciferase reporter in HEK293T cells. EC50 = 50 pg/ml. Bioactivity is determined using a TGF-β3 responsive firefly luciferase reporter in HEK293T cells. Cells are treated (in triplicate) with a serial dilution of TGF-β3 for 6 hours. Firefly luciferase activity is measured and normalised to the control Renilla luciferase activity. Data from Qk054 batch #104369.

TGF-β3 migrates as a single band at 25 kDa in non-reducing (NR) conditions and 13 kDa upon reduction (R).  No contaminating protein bands are visible. 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 Qk054 batch #104369.

Protein background

Transforming growth factor beta 3 (TGF-β3) is a member of the TGF-β family (and TGF-β superfamily). Like other TGF-β family members – TGF-β1 and 2 – TGF-β3 signals via receptor complexes consisting of Type I and Type II serine/threonine kinase receptors, activating intracellular SMAD transcription factors [1]. The TGF-β family have a high degree of overlap/redundancy and regulate a variety of cellular functions, including cell survival, proliferation, and differentiation [2]. TGF-β3 has isoform-specific roles in embryonic palate fusion [3] and wound healing [4].

In culture, TGF-β3 is often used in human pluripotent cell maintenance. TGF-β3 is a component of B8 media – the low-cost weekend-free iPSC media developed in the lab of Paul Burridge, Northwestern University [5]. In the development of B8 media, Kuo et al. proved that while TGF-β1 can maintain iPSCs at 2 ng/ml, TGF-β3 can maintain iPSCs in as little as 0.1 ng/ml, making it a cost-effective alternative to TGF-β1 [5].

Background references

1. T. Huang, S. L. Schor and A. P. Hinck. Biological Activity Differences between TGF-β1 and TGF-β3 Correlate with Differences in the Rigidity and Arrangement of Their Component Monomers. Biochemistry 2014, 53, 36, 5737–5749. doi.org/10.1021/bi500647d
2. M. Morikawa, R. Derynck and K. Miyazono. TGF-β and the TGF-β Family: Context-Dependent Roles in Cell and Tissue Physiology. Cold Spring Harb Perspect Biol. 2016 May 2;8(5):a021873. doi: 10.1101/cshperspect.a021873
3. G, Proetzel, S. A. Pawlowski, M. V. Wiles, M. Yin, G. P. Boivin, P. N. Howles, J. Ding, M. W. Ferguson and T. Doetschman. Transforming growth factor-beta 3 is required for secondary palate fusion. Nat Genet. 1995 Dec;11(4):409-14. doi: 10.1038/ng1295-409
4. B. Bandyopadhyay, J. Fan, S. Guan, Y. Li, M. Chen, D. T. Woodley, W and Li. A “traffic control” role for TGFbeta3: orchestrating dermal and epidermal cell motility during wound healing. J Cell Biol. 2006 Mar 27;172(7):1093-105. doi: 10.1083/jcb.200507111
5. H. H. Kuo, X. Gao, J. M. DeKeyser, K. A. Fetterman, E. A. Pinheiro, C. J. Weddle, H. Fonoudi, M. V. Orman, M. Romero-Tejeda, M. Jouni, M. Blancard, T. Magdy, C. L. Epting, A. L. George Jr and P. W. Burridge. Negligible-Cost and Weekend-Free Chemically Defined Human iPSC Culture. Stem Cell Reports. 2020 Feb 11;14(2):256-270. doi: 10.1016/j.stemcr.2019.12.007

Publications using recombinant human TGF-β3 protein (Qk054)

Nutritional requirements of human induced pluripotent stem cells
Lyra-Leite DM, Copley RR, Freeman PP et al.
DOI: 10.1016/j.stemcr.2023.05.004

Refined home-brew media for cost-effective, weekend-free hiPSC culture and genetic engineering
Truszkowski L, Bottini S, Bianchi S et al.
DOI: https://doi.org/10.12688/openreseurope.18245.1

Thalamocortical organoids enable in vitro modeling of 22q11.2 microdeletion associated with neuropsychiatric disorders
Shin D, Kim CN, Ross J et al.
DOI: 10.1016/j.stem.2024.01.010

FAQ

What is TGF beta 3?
TGF-β3 is a growth factor involved in tissue development, wound healing, and immune regulation, crucial for iPSC and ESC proliferation and differentiation.

Where is TGF-β3 found?
TGF-β3 is found in various tissues throughout the body, including the skin, bones, blood vessels, and immune cells.

Is TGF-β3 a cytokine?
Yes, TGF-β3 is a cytokine.

What does the TGF-β3 gene do?
The TGF-β3 gene encodes the transforming growth factor beta 3 protein.

What does TGF-β3 bind to?
TGF-β3 binds to TGF-β receptors. This triggers intracellular signaling pathways that regulate gene expression, influencing cellular processes.

What is the function of the TGF-β3 receptor?
The TGF-β3 receptor serves as a cell surface receptor for the TGF-β3 ligand. It plays a crucial role in transmitting signals from the extracellular environment to the interior of the cell upon ligand binding and initiating intracellular signaling cascades that regulate gene expression.

What is the TGF-β3 pathway?
The TGF-β3 pathway is a series of intracellular signaling events triggered by the binding of TGF-β3 ligands to their cell surface receptors. Upon ligand binding, the TGF-β3 receptors undergo phosphorylation and activation, leading to the activation of downstream signaling molecules such as Smad proteins. These Smad proteins translocate into the nucleus, where they regulate the expression of target genes involved in various cellular processes such as proliferation and differentiation.

How is TGF-β3 used in cell culture?
TGF-β3 regulates cell behavior, promoting differentiation and wound healing. It is often used in tissue engineering, cellular agriculture industries for cultivated meat, and immune modulation.