FGF4 – human (Qk004)

Fibroblast growth factor 4 (FGF-4) is a member of the FGF superfamily with a physiological role in the regulation of proliferation and differentiation in embryonic stem cells and tissue stem cells 1–3

FGF-4 has been shown to promote neural stem cell proliferation and neuronal differentiation in the postnatal brain 4, increase the proliferation rate of human adult bone-marrow derived mesenchymal stem cells 5 and support the maintenance, proliferation and self-renewal properties of human embryonic stem cells 6

Synergism between FGF4 and WNT signalling acts to form hindgut organoids from iPSC-derived human posterior gut endoderm cells7 and in FGF4 can be used in culture systems to mimic embryonic intestinal development during directed differentiation of pluripotent stem cells into intestinal organoids

  1. Beenken, A. & Mohammadi, M. The FGF family: biology, pathophysiology and therapy. Nat. Rev. Drug Discov. 8, 235–53 (2009).
  2. Coutu, D. L. & Galipeau, J. Roles of FGF signaling in stem cell self-renewal, senescence and aging. Aging (Albany. NY). 3, 920–33 (2011).
  3. Kosaka, N., Sakamoto, H., Terada, M. & Ochiya, T. Pleiotropic function of FGF-4: Its role in development and stem cells. Dev. Dyn. 238, 265–276 (2009).
  4. Kosaka, N. et al. FGF-4 regulates neural progenitor cell proliferation and neuronal differentiation. FASEB J. 20, 1484–1485 (2006).
  5. Farré, J. et al. FGF-4 increases in vitro expansion rate of human adult bone marrow-derived mesenchymal stem cells. Growth Factors 25, 71–76 (2007).
  6. Mayshar, Y. et al. Fibroblast Growth Factor 4 and Its Novel Splice Isoform Have Opposing Effects on the Maintenance of Human Embryonic Stem Cell Self-Renewal. Stem Cells 26, 767–774 (2008).
  7. Zhang, R.-R. et al. Stem Cell Reports Human iPSC-Derived Posterior Gut Progenitors Are Expandable and Capable of Forming Gut and Liver Organoids. Stem Cell Reports 10, 780–793 (2018).
  8. McCracken, K. W. et al. Modelling human development and disease in pluripotent stem-cell-derived gastric organoids. Nature 516, 400–404 (2014).
Fibroblast Growth Factor-4, Kaposi’s sarcoma-associated FGF, k-FGF, Heparin secretory-transforming protein 1 HST-1, Transforming protein KS3, Heparin-binding growth factor 4, HBGF-4

Summary: Mature domain of human FGF4 (residues 79-206, Uniprot:P08620) expressed in E.coli and purified to homogeneity. Mature protein is a non-glycosylated protein with a molecular weight of ca. 14 kDa

Form: protein is provided frozen in PBS (carrier protein-free) at 1 mg/ml

Molecular mass: ~14 kDa

Thaw the sample on ice, spin briefly and dilute with PBS as needed.  Our protein are supplied carrier-protein free.  If compatible with your work, add carrier protein of your choice such as BSA, HSA or gelatin to further minimise 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.

Result: FGF4 migrates as a single band at 14 kDa in non-reducing (NR) and upon reduction (R).  No contaminating protein bands are visible.

Purified recombinant 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.

Result: FGF4 induces cell proliferation of HEK293T cells.

Human embryonic kidney cell line 293T (HEK293T) was cultured in serum-free media containing 100ng/ml hFGF2. The number of viable metabolically active cells was detected by ATP measurement using a bioluminescent reaction (n=3; data show mean ± SEM. Data provided by Stemnovate Ltd, Cambridge, UK.

Result: FGF4 induces cell proliferation of iPSC cells (Stemnovate iPSC1 line).

IPSC (Stemnovate iPSC1) were cultured in serum-free media containing 100ng/ml hFGF4. The number of viable metabolically active cells was detected by ATP measurement using a bioluminescent reaction (n=3; data show mean ± SEM). No media change was performed on day 2 leading to reduced viable cell numbers on day 3. Data provided by Stemnovate Ltd, Cambridge, UK.

Result: calculated molecular mass of the FGF4 is 14409.8 Da. Result of the analysis: 14278.4 Da which is consistent with the calculated mass.  No significant heterogeneity is present.

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 disulphide-linked. The different peaks represent different charge states of the protein.

Result: UV spectrum shows full recovery of protein following aliquoting and lyophilisation.

Absorbance at 280 nm: average 0.068
Recovered concentration: 0.068 cm-1 x 10 / 0.62 cm-1 mg ml-1 = 1.1 mg / ml
Recovery: 110% (>100% due to routine 10% 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

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 optimise 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.

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.

View full batch quality testing data for Qk001

Batch #010

All our proteins are produced in-house by our scientists and we provide detailed quality data for each individual batch.  Please contact us any time by email support@qkine.com or phone +44 (0) 1223 491486 if you have any questions.

After aliquotting and lyophilising the protein, we choose a vial at random and reconstitute as recommended to ensure we are testing as close to the protein you will receive as possible.  Biochemical identity and purity of each batch is checked using SDS-PAGE, mass spectrometry and analytical reverse phase chromatography.  We use a sensitive test to ensure endotoxin levels are at industry leading low levels (<0.01 EU per µg protein). Bioactivity of the protein is determined using the quantitative activin responsive luciferase reporter assay.  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.

View units sizes available and purchasing information online or email orders@qkine.com

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