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| 1mg | ||
| 5mg | ||
| 10mg | ||
| Other Sizes |
| Targets |
EGF binds with high affinity to the epidermal growth factor receptor (EGFR, also known as ErbB1/HER1), a transmembrane receptor tyrosine kinase. Upon ligand binding, EGFR undergoes dimerization, autophosphorylation at specific tyrosine residues, and activation of downstream signaling cascades including the MAPK/ERK pathway, PI3K/Akt pathway, and PLCgamma/PKC pathway, leading to cell proliferation, survival, migration, and differentiation.
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| ln Vitro |
In wound healing applications, epidermal growth factor (EGF) is used [1]. In the fibroblast BALB/c3T3 cell line, epidermal growth factor (EGF) promotes cell division. In addition to inducing EGF receptor expression and eliciting proliferating cell nuclear antigen, the hydrogel's released epidermal growth factor (EGF) retained its biological activity and demonstrated therapeutic potential to improve diabetic wound healing.
In vitro, EGF is a potent mitogen that stimulates the proliferation of fibroblasts (BALB/c3T3 cell line), epithelial cells, and endothelial cells. EGF released from hydrogels maintains its bioactivity, induces EGF receptor expression and proliferating cell nuclear antigen (PCNA), and promotes cell migration. It initiates a wide variety of cellular events ranging from early events (e.g., stimulation of amino acid or ion transport within minutes) to later events such as commitment of cells to DNA synthesis. EGF also stimulates the phosphorylation of pyruvate kinase in isolated rat hepatocytes and gluconeogenesis. |
| ln Vivo |
In vivo, EGF accelerates wound healing, promotes epithelialization and granulation tissue formation, and enhances diabetic wound healing in animal models. It stimulates the proliferation of epidermal cells and is used in wound healing applications. EGF also promotes angiogenesis and vascular formation. It has been studied in diabetic models where EGF released from hydrogels shows enhanced therapeutic potential for diabetic wound healing. EGF is also involved in the development of various tissues and organs during embryogenesis.
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| Enzyme Assay |
For receptor binding assays, immobilize purified EGFR (0.5-1 ug/well) on 96-well plates. Block with 1% BSA in PBS for 1 hour at room temperature. Add 125I-labeled EGF (10-100 pM) with or without unlabeled EGF or EGF phosphate (1 pM-1 uM) in binding buffer (50 mM HEPES, 0.1% BSA, pH 7.4). Incubate for 2 hours at 4degC. Wash 3 times with cold PBS, add scintillation fluid, and measure bound radioactivity in a gamma counter. For non-radioactive assays, use FITC-labeled EGF and measure fluorescence polarization. Calculate binding affinity (Kd) and competitive IC50 values by nonlinear regression.
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| Cell Assay |
Culture BALB/c3T3 fibroblasts or human keratinocytes in DMEM with 10% FBS at 37degC with 5% CO2. For proliferation assays, serum-starve cells for 12-24 hours in 0.5% FBS medium. Treat with EGF phosphate at concentrations of 0.1-100 ng/mL for 24-48 hours. Assess cell proliferation by MTT assay, cell counting, or 3H-thymidine incorporation. For signaling studies, treat cells with EGF (10-100 ng/mL) for 0-60 minutes. Prepare cell lysates and perform Western blot analysis using antibodies specific for phospho-EGFR (Tyr1068), phospho-ERK1/2 (Thr202/Tyr204), phospho-Akt (Ser473), and downstream targets. For wound healing scratch assays, create a scratch in confluent monolayers, wash with PBS, add medium with EGF (10-50 ng/mL), and monitor wound closure by microscopy at 0, 6, 12, and 24 hours. For cell migration assays, use Transwell chambers with 8 um pore size. Add EGF to the lower chamber (0.1-100 ng/mL) as a chemoattractant.
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| Animal Protocol |
For wound healing studies, use diabetic rat models (e.g., STZ-induced diabetic rats) or db/db mice. Create full-thickness excisional wounds (6-8 mm diameter) on the dorsum. Apply EGF phosphate in hydrogel or solution topically once daily for 7-14 days. Monitor wound closure by digital photography and planimetry. At sacrifice, excise wound tissues, fix in formalin, embed in paraffin, section, and stain with H&E for histological evaluation of re-epithelialization and granulation tissue formation. Perform immunohistochemistry for proliferating cell nuclear antigen (PCNA), Ki-67, and CD31 (angiogenesis). For non-diabetic wound models, use normal rats or mice. Quantify wound healing rate as percentage of original wound area.
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| ADME/Pharmacokinetics |
EGF is a 53-amino acid peptide with a molecular weight of approximately 6.2 kDa. Following systemic administration (intravenous or subcutaneous), EGF has a short plasma half-life in rodents of 10-30 minutes due to rapid clearance by receptor-mediated endocytosis and proteolytic degradation in the liver and kidneys. The phosphate salt form may enhance formulation stability without significantly altering pharmacokinetics. Topical application results in minimal systemic absorption, with the majority of EGF remaining localized to the wound site. Tear EGF concentrations rapidly increase and return to baseline after 4 hours following topical eye drop administration in humans, with no detectable change in serum EGF levels.
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| Toxicity/Toxicokinetics |
EGF is generally well-tolerated at therapeutic doses for topical wound healing applications. No significant systemic toxicity has been observed in animal studies or human clinical trials with topical EGF. Recombinant human EGF (rhEGF) eye drops have been evaluated in Phase I clinical trials in healthy subjects, demonstrating good safety and tolerability with no serious adverse events. Systemic administration of high doses of EGF may theoretically promote cell proliferation, including potential neoplastic growth, but this is not a concern for topical wound healing applications. Standard laboratory precautions for handling proteins (avoid inhalation, skin contact) should be followed.
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| References |
[1]. Wong WR, et al. Applications, and efficient large-scale production, of recombinant human epidermal growth factor. Biotechnol Genet Eng Rev. 2001;18:51-71.
[2]. Lao G, et al. Controlled release of epidermal growth factor from hydrogels accelerates wound healing in diabetic rats. J Am Podiatr Med Assoc. 2012 Mar-Apr;102(2):89-98. |
| Additional Infomation |
EGF phosphate is a recombinant human epidermal growth factor (rhEGF) peptide produced by recombinant DNA technology. It is used in research applications including wound healing, tissue engineering, cell culture (as a supplement for stem cell, organoid, and epithelial cell culture), and studies of cell proliferation and invasion. The phosphate formulation enhances stability in aqueous solutions. For research use only; not for clinical therapy without regulatory approval.
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| Molecular Weight |
6-12KDa
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| Related CAS # |
Epidermal growth factor (EGF);62253-63-8
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| Appearance |
Typically exists as solid at room temperature
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| HS Tariff Code |
2934.99.9001
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| Storage |
Powder -20°C 3 years 4°C 2 years In solvent -80°C 6 months -20°C 1 month Note: Please store this product in a sealed and protected environment (e.g. under nitrogen), avoid exposure to moisture and light. |
| Shipping Condition |
Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)
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| Solubility (In Vitro) |
H2O :≥ 3.33 mg/mL
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| Solubility (In Vivo) |
Note: Listed below are some common formulations that may be used to formulate products with low water solubility (e.g. < 1 mg/mL), you may test these formulations using a minute amount of products to avoid loss of samples.
Injection Formulations
Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline)(e.g. IP/IV/IM/SC) *Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH ₂ O to obtain a clear solution. Injection Formulation 2: DMSO : PEG300 ：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL DMSO → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline) Injection Formulation 3: DMSO : Corn oil = 10 : 90 (i.e. 100 μL DMSO → 900 μL Corn oil) Example: Take the Injection Formulation 3 (DMSO : Corn oil = 10 : 90) as an example, if 1 mL of 2.5 mg/mL working solution is to be prepared, you can take 100 μL 25 mg/mL DMSO stock solution and add to 900 μL corn oil, mix well to obtain a clear or suspension solution (2.5 mg/mL, ready for use in animals). View More
Injection Formulation 4: DMSO : 20% SBE-β-CD in saline = 10 : 90 [i.e. 100 μL DMSO → 900 μL (20% SBE-β-CD in saline)] Oral Formulations
Oral Formulation 1: Suspend in 0.5% CMC Na (carboxymethylcellulose sodium) Oral Formulation 2: Suspend in 0.5% Carboxymethyl cellulose Example: Take the Oral Formulation 1 (Suspend in 0.5% CMC Na) as an example, if 100 mL of 2.5 mg/mL working solution is to be prepared, you can first prepare 0.5% CMC Na solution by measuring 0.5 g CMC Na and dissolve it in 100 mL ddH2O to obtain a clear solution; then add 250 mg of the product to 100 mL 0.5% CMC Na solution, to make the suspension solution (2.5 mg/mL, ready for use in animals). View More
Oral Formulation 3: Dissolved in PEG400 (Please use freshly prepared in vivo formulations for optimal results.) |
Calculation results
Working concentration： mg/mL；
Method for preparing DMSO stock solution： mg drug pre-dissolved in μL DMSO (stock solution concentration mg/mL). Please contact us first if the concentration exceeds the DMSO solubility of the batch of drug.
Method for preparing in vivo formulation:：Take μL DMSO stock solution, next add μL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL ddH2O，mix and clarify.
(1) Please be sure that the solution is clear before the addition of next solvent. Dissolution methods like vortex, ultrasound or warming and heat may be used to aid dissolving.
(2) Be sure to add the solvent(s) in order.
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