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Recombinant human EGF protein (Qk011)
Human epidermal growth factor (EGF) protein is a potent EGF-family growth factor used in many human and mouse organoid and stem cell culture systems including intestinal and tumor organoid culture. EGF is also used in epithelial cell culture.
Qkine human EGF (Qk011) is a highly pure, animal origin-free, bioactive 6.3 kDa recombinant protein, making it ideal for use in chemically defined induced pluripotent stem cell (iPSC), embryonic stem cell (ESC) and organoid culture media.
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1000µg will be despatched as 2 x 500µg
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Summary
High purity human EGF protein (residues 971-1023, Uniprot: P01133)
>98%, by SDS-PAGE quantitative densitometry
6.3 kDa
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 at >100 µg/ml (provided with protein and free of charge), prepare single use aliquots, add carrier protein if desired and store frozen at -20°C or -80°C
Featured applications
Tumor organoid culture
Human
Species similarity:
Porcine – 85%
Mouse – 74%
Rat – 70%
Bioactivity
EGF activity is determined using the Promega serum response element luciferase reporter assay (*) in transfected HEK293T cells. Cells are treated in triplicate with a serial dilution of EGF for 3 hours. Firefly luciferase activity is measured and normalized to the control Renilla luciferase activity. EC50 = 11.6 pM (72.8 pg/mL). Data from Qk011 lot #10. *Promega pGL4.33[luc2P/SRE/Hygro] #E1340
Purity
EGF protein migrates as a single band at ~12 kDa in non-reducing (NR) and 9 kDa in reducing (R) conditions. 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. Data from Qk011 lot #011.
Further quality assays
Mass spectrometry: single species with expected mass
Endotoxin: <0.005 EU/μg protein (below level of detection)
Recovery from stock vial: >95%
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.
Qkine EGF is as biologically active as a comparable alternative supplier protein
Quantitative luciferase assay with Qkine EGF (Qk011, green) and alternative supplier EGF (Supplier B, black). Cells were treated in triplicate with a serial dilution of EGF for 3 hours. Firefly luciferase activity was measured and normalized to control Renilla luciferase activity. EC50s 0.11 ng/ml.
Qkine EGF remains stable and bioactive in conditioned media
Quantitative luciferase assay with Qkine EGF (Day 0, green) and (Day 2, black). Cells were treated in triplicate with a serial dilution of EGF for 3 hours after preincubation in conditioned media. Firefly luciferase activity was measured and normalized to control Renilla luciferase activity.
Protein background
Members of the human epidermal growth factor family are synthesized as type I transmembrane precursor proteins, often containing several EGF domains in the extracellular region. Proteolysis yields mature proteins that are released from the cell surface [1]. Epidermal growth factor binds to high-affinity EGF receptors (EGFRs) and promotes receptor dimerization and clustering leading to activation of downstream signaling pathways, including PI3K, ERK1/2, JAK/STAT, β-catenin, and calcium signaling [2].
Human epidermal growth factor (EGF) stimulates cell proliferation and differentiation. It is used extensively in induced-pluripotent stem cell (iPSC) and embryonic stem cell (ESC) culture systems for the successful expansion and differentiation of epithelial, neural, mesoderm and hematopoietic lineages [3]. EGF also stimulates the differentiation of mesenchymal stem cells (MSC) into bone-forming cells [4].
Recombinant EGF protein is a key component of many organoid media, along with other stem cell niche factors, R-Spondin 1, Noggin or Gremlin, Wnt3a and FGF10. In addition, recombinant EGF is used for establishing and maintaining intestine, stomach, liver, pancreas, brain and cancer organoids [5].
Publications using recombinant human EGF protein (Qk011)
Generation of human iPSC-derived pancreatic organoids to study pancreas development and disease
bioRxiv preprint on 25 October 2024 by Darrigrand, J. et al.
Metabolic rewiring underpins human trophoblast induction.
Preprint on 18 March 2024 by Żylicz, J., van Nerum, K., Wenzel, A., Argemi Muntadas, L. et al.
FAQ
Epidermal growth factor (EGF) is a growth factor which stimulates cell proliferation and differentiation.
Originally found in salivary glands and urine, EGF is known to be expressed in many human tissues, including platelets.
EGF primarily stimulates growth of epidermal and epithelial cells, but is also used for expansion of stem cells in feeder-free culture systems.
Epidermal growth factor receptor (EGFR) family are receptor tyrosine kinase (RTK) proteins. There are 4 members: EGFR (ErbB1, HER1), ErbB2 (HER2), ErbB3 (HER3) and ErbB4 (HER4).
Tyrosine phosphorylation of EGF receptors stimulates various signaling pathways, including Ras/MAPK, PLCγ1/PKC, PI3 kinase/Akt, and STAT pathways.
EGF can be added to many stem cell cultures to maintain them in their pluripotent state.
Our products are for research use only and not for diagnostic or therapeutic use. Products are not for resale.
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