Qk003 Recombinant human FGF10 protein is a 17 kDa non-glycosylated, bioactive domain of human fibroblast growth factor 10. Our recombinant FGF10 protein has been optimized for expression in E. coli and purified to homogeneity for use in ESC/iPSC maintenance and differentiation, and for the study of epithelial to mesenchymal transition and tumor metastasis.
Recombinant human FGF10 protein
£160.00 – £1,200.00
- Description
- Purity & bioactivity data
- Customer & collaborator data
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- Resuspension & storage
Qk003 Recombinant human FGF10 protein
Fibroblast growth factor 10 (FGF10) is involved in a number of different embryo and adult cell and tissue types, including mesenchymal, neuronal and epithelial cells. Human FGF10 protein is expressed in the mesenchyme and functions through interacting with the epithelial FGF Receptor 2b (Fgfr2b)1. It has also been shown to interact weakly with FGF Receptor 1b2. The mature form of human FGF10 protein is an approximately 20 kDa protein highly similar to FGF7 and with a serine-rich region near its N-terminus3. It is secreted by mesenchymal cells and is bound and activated by extracellular FGF-BP4.
Human fibroblast growth factor 10 is first active in the limb bud mesoderm where it creates and maintains FGF signalling with epithelial FGF8, then drives a positive feedback loop accumulating mesenchyme in the growing bud, and finally induces the apical ectodermal ridge which ultimately gives rise to feet and hands5. Lung development is based on the same epithelial-mesenchymal FGF mediations involving FGF10 from the foregut mesenchyme signalling to FGFR2 in the foregut epithelium6.
Furthermore, FGF10 protein is involved in the development of white adipose tissue, heart, liver, brain, kidney, thymus, inner ear, tongue, trachea, eye, prostate, salivary gland and mammary gland. It has been shown to induce migration and invasion of pancreatic cancer cells and to be associated with breast cancer risk, and patients with FGF10 haploinsufficiency present symptoms of chronic obstructive pulmonary disease. Human recombinant FGF10 protein also drives the differentiation of embryonic stem cells into gut-like structures, cardiomyocytes and hepatocytes1.
Summary: Mature domain of human FGF10 protein (residues 64-208, Uniprot: O15520) expressed in E. coli and purified to homogeneity. Mature protein is a non-glycosylated protein with a molecular mass of 17 kDa
Molecular mass: ~17 kDa
Form: Protein is provided frozen in PBS (carrier protein-free). Protein concentration is 1 mg/ml.
Fibroblast Growth Factor 10, FGFA, KGF2, Keratinocyte growth factor 2
- Itoh, N. & Ohta, H. Fgf10: A Paracrine-Signaling Molecule in Development, Disease, and Regenerative Medicine. Curr. Mol. Med. 14, 504–509 (2014).
- Zhang, X. et al. Receptor specificity of the fibroblast growth factor family. The complete mammalian FGF family. J. Biol. Chem. 281, 15694–700 (2006).
- Igarashi, M., Finch, P. W. & Aaronson, S. A. Characterization of recombinant human fibroblast growth factor (FGF)-10 reveals functional similarities with keratinocyte growth factor (FGF-7). J. Biol. Chem. 273, 13230–5 (1998).
- Beer, H.-D. et al. The fibroblast growth factor binding protein is a novel interaction partner of FGF-7, FGF-10 and FGF-22 and regulates FGF activity: implications for epithelial repair. Oncogene 24, 5269–77 (2005).
- Ohuchi, H. et al. The mesenchymal factor, FGF10, initiates and maintains the outgrowth of the chick limb bud through interaction with FGF8, an apical ectodermal factor. Development 124, 2235–44 (1997).
- Min, H. et al. Fgf-10 is required for both limb and lung development and exhibits striking functional similarity to Drosophila branchless. Genes Dev. 12, 3156–61 (1998).
- Zhang, R.-R. et al. Human iPSC-Derived Posterior Gut Progenitors Are Expandable and Capable of Forming Gut and Liver Organoids. Stem Cell Reports 10, 780–793 (2018).
- McCracken, K. W. et al. Modelling human development and disease in pluripotent stem-cell-derived gastric organoids. Nature 516, 400–404 (2014).
Purity and bioactivity
For peace of mind that our recombinant human FGF10 protein will work exactly the same way every day, from batch to batch and at any scale you need, we conduct extensive purity and bioactivity analysis, including SDS-PAGE, mass spectrometry, reversed-phase chromatography, UV spectroscopy and endotoxin level testing.
We also work with our collaborators at Stemnovate, a specialist stem cell biotechnology company, to check the final purified recombinant FGF10 protein supports proliferation and promotes epithelial to mesenchymal transition in human primary keratinocytes
Find out more about our extensive purity testing in the next tab.
Example data
Protein purity: SDS-PAGE in reduced and non-reduced conditions
Bioactivity: proliferation and EMT (Stemnovate data)
Research applications
iPSC/ESC differentiation
Organoid
All our proteins are produced in our Cambridge, UK labs. We provide detailed quality data for each batch because we believe reliable, high quality cytokines and growth factors are critical for successful stem cell and organoid culture. Please contact us with questions any time by email at support@qkine.com or phone +44 (0) 1223 491486 / US toll free 1-866 877 2185.
Order online and upload your PO or pay by credit card, or email your PO to orders@qkine.com.
We provide bulk orders and stock reservation for sensitive applications, please email us.
Our products are for research use only and not for diagnostic or therapeutic use. Products are not for resale.
For final purity and activity tests on our proteins, we choose a vial at random and reconstitute as recommended. Biochemical identity and purity is determined using SDS-PAGE, mass spectrometry and analytical reversed-phase chromatography. Bioactivity is quantified using an appropriate cell-based assay. As stem cells are sensitive to endotoxin levels, we use a high resolution test to ensure endotoxin levels are at industry-leading low levels (<0.01 EU per µg protein). We also check that the correct amount of protein is recovered from the vial - it might sound basic but if you order 100 µg, we believe you should receive 100 µg, so when you use the proteins you can rely on your calculated dilution.
Purity: SDS-PAGE
Result: FGF10 migrates as a single band at 17 kDa in non-reducing (NR) conditions and upon reduction (R). No contaminating protein bands are visible.
Purified recombinant human FGF10 protein (7 µg) was resolved using 15% w/v SDS-PAGE in reduced (+β-mercaptothanol, R) and non-reduced conditions (NR) and stained with Coomassie Brilliant Blue R250.
Bioactivity: Induction of EMT in primary keratinocytes
Result: FGF10 supports proliferation and promotes epithelial to mesenchymal transition in human primary keratinocytes. Data and evaluation by Stemnovate Ltd.
Epithelial to mesenchymal transition (EMT) is a crucial morphogenetic process during development in which cells lose their epithelial characteristics and acquire migratory mesenchymal properties. FGF10 has an important role both during the embryonic EMT (type I) and on cancer cell initiation of metastasis (type III EMT).
Cell proliferation assays to assess the effect of Qkine FGF10 (0-100 ng/ml) on human primary epidermal keratinocytes in serum-free keratinocyte media. Cells were evaluated at culture days: 0 (baseline), 1, 2, 3, 4 days, as summarized schematically in Figure 1a. Figure 1b shows cell proliferation (Relative Luminescence Unit [RFU]) for days 1, 2, 3, 4 and normalized to day 0 readouts (n=3; P*<0.05 vs control). The log concentration plot in Figure 1c shows percent cell proliferation normalized over untreated control (%) and to day 0 (baseline) after 4 days treatment (n=3; P*<0.05). The maximal cell proliferation was observed at ~10ng/ml FGF10 and a reduction in cell number/viability as observed at 100 ng/ml. Data provided by Stemnovate Ltd, Cambridge, UK.
Purity: mass spec analysis
Result: calculated molecular mass of the FGF10 is 16912.4 Da. Result of the analysis: 16781.2 Da which is consistent with the calculated mass. There are no minor contaminants.
MALDI mass spectrometric analysis is used to confirm the molecular mass of the intact protein and to reveal any heterogeneity that would not be evident in SDS-PAGE analysis. The results are compared with calculated mass of the protein with the assumption that all the cysteines are disulfide-linked. The different peaks represent different charge states of the protein.
Purity: Endotoxin level determination
Result: Endotoxin level <0.005 EU/ug protein (below level of detection)
Stem cell cultures are sensitive to endotoxins1, which can be present in media, serum and as a contaminant on plasticware. We optimize our protein production processes to ensure the lowest possible levels of endotoxin contamination. Our endotoxin pass criteria are set at the industry-leading <0.1 EU per ug protein and we aim for <0.01 EU per ug protein. Endotoxin levels in our proteins are determined by an external expert microbiological testing services provider. Example data from Qk001 batch #011.
1. A biological study establishing the endotoxin limit for in vitro proliferation of human mesenchymal stem cells (2017). Yusuke Nomura, Chie Fukui, Yuki Morishita, Yuji Haishima. Regenerative Therapy, 7, 45-51.
Result: FGF10 supports epithelial to mesenchymal transition (EMT) in human primary keratinocytes. Data and evaluation by Stemnovate Ltd.
Epithelial to mesenchymal transition (EMT) is a crucial morphogenetic process during development in which cells lose their epithelial characteristics and acquire migratory mesenchymal properties. Human FGF10 protein has an important role both during the embryonic EMT (type I) and on cancer cell initiation of metastasis (type III EMT).
Induction of EMT in human primary keratinocytes following treatment with hFGF10. Induction of EMT was evaluated using immunofluorescence staining to determine expression of the epithelial marker (Cytokeratin 14 [CK14]) and mesenchymal marker (α-Smooth Muscle Actin [αSMA]) in human primary epidermal keratinocytes after 4 days treatment with Qk003 hFGF10 (0-100 ng/ml).
Thaw the sample on ice, spin briefly and dilute with PBS as needed. Our proteins are supplied carrier protein-free. If compatible with your work, add a carrier protein of your choice such as BSA, HSA or gelatin to further minimize loss by adsorption. Spin in a microfuge for 5 minutes at maximum speed and divide the solution into suitable aliquots and store at -80°C. We recommend that single-use aliquots should be prepared to avoid freeze-thaw cycles.
Every effort is made to ensure samples are sterile; however, we recommend sterile filtering after dilution in media or the final working solution.
We check that the correct amount of protein is recovered from the vial – it might sound basic but if you order 100 µg, we believe you should receive 100 µg, so you can rely on your calculated dilution.
Recovery: protein quantitation
Result: UV spectrum shows full recovery of protein following aliquoting and lyophilization.
Absorbance at 280 nm: average 0.172
Recovered concentration: 0.172 cm-1 x 10 / 0.144 cm-1 mg ml-1 = 1.2 mg / ml
Recovery: 120% (>100% due to routine 10-20% over-fill of vials during aliquoting)
The sample was diluted 1:10 in 100 mM sodium phosphate pH 7.4 and the UV spectrum 340-220 nm measured in duplicate. Concentration was calculated using extinction coefficient at 280 nm.
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