E. coli-derived human EGF protein Asn971-Arg1023, with an N-terminal Met Produced using non-animal reagents in an animal-free laboratory. Manufactured and tested under cGMP guidelines.
>97%, by SDS-PAGE under reducing conditions and visualized by silver stain. The result of the major peak from mass spectrometry analysis is 6347 Da, which corresponds to the calculated molecular mass of 6353 Da.
<0.10 EU per 1 μg of the protein by the LAL method.
N-terminal sequence Analysis
Predicted Molecular Mass
6 kDa, reducing conditions
Measured in a cell proliferation assay using Balb/3T3 mouse embryonic fibroblast cells. Rubin, J.S. et al. (1991) Proc. Natl. Acad. Sci. USA 88:415. The ED50 for this effect is 20‑100 pg/mL. The specific activity of Recombinant Human EGF is >8.0 x 105 IU/mg, which is calibrated against the human EGF WHO International Standard (NIBSC code: 91/530).
Recombinant Human EGF GMP Protein, CF Scientific Data Examples
GMP-grade Recombinant Human EGF (Catalog # 236‑GMP) stimulates proliferation of the Balb/3T3 mouse embryonic fibroblast cell line. The ED50for this effect is 20‑100 pg/mL. Three independent lots were tested for activity and plotted on the same graph to show lot-to-lot consistency of GMP EGF.
1 µg/lane of GMP-grade Recombinant Human EGF (Catalog # 236-GMP) was resolved by SDS-PAGE and visualized by silver staining under reducing (R) conditions, showing a single band at 6 kDa.
MALDI‑TOF analysis of GMP‑grade Recombinant Human EGF (Catalog # 236‑GMP). The major peak corresponds to the calculated molecular mass, 6353 Da. The minor peak at 6549 Da is a matrix-associated artifact of the MALDI-TOF.
Formulation, Preparation and Storage
What does CF mean?
CF stands for Carrier Free (CF). We typically add Bovine Serum Albumin (BSA) as a carrier protein to our
Adding a carrier protein enhances protein stability, increases shelf-life, and allows the recombinant
protein to be stored at a more dilute concentration.
The carrier free version does not contain BSA.
What formulation is right for me?
In general, we advise purchasing the recombinant protein with BSA for use in cell or tissue culture, or
as an ELISA standard.
In contrast, the carrier free protein is recommended for applications, in which the presence of BSA
Lyophilized from a 0.2 μm filtered solution in PBS.
Reconstitute at 200 μg/mL in PBS.
The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage:
Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
A minimum of 12 months when stored at ≤ -20 °C as supplied. Refer to lot specific COA for the Use by Date.
1 month, 2 to 8 °C under sterile conditions after reconstitution.
3 months, ≤ -20 °C under sterile conditions after reconstitution.
Epidermal growth factor (EGF) is a small, potent growth factor capable of inducing cell proliferation, differentiation, and survival. EGF is the founding member of the EGF family that also includes TGF-alpha, amphiregulin (AR), betacellulin (BTC), epiregulin (EPR), heparin‑binding EGF‑like growth factor (HB‑EGF), epigen, and the neuregulins (NRG)-1 through -6 (1). Members of The EGF family are characterized by a shared structural motif, the EGF‑like domain, which contains three intramolecular disulfide bonds that are formed by six similarly spaced, conserved cysteine residues (2). These disulfide bonds are essential for proper protein conformation and receptor binding. All EGF family members are synthesized as type I transmembrane precursor proteins that may contain several EGF domains in the extracellular region. The mature proteins are released from the cell surface by regulated proteolysis (1). The full length EGF protein is 1207 amino acids (aa) (EGF precursor) containing nine EGF domains and nine LDLR class B repeats. However, the mature protein is much smaller, only 53 aa, and is generated by proteolytic cleavage of the EGF domain proximal to the transmembrane region (3). EGF is well conserved across mammals with mature human EGF 70% identical to mature mouse and rat EGF. Physiologically, EGF is found in various body fluids, including blood, milk, urine, saliva, seminal fluid, pancreatic juice, cerebrospinal fluid, and amniotic fluid (4). EGF is a high affinity ligand of the EGF receptor (ErbB). Four ErbB (HER) family receptor tyrosine kinases including EGFR/ErbB1, ErbB2, ErbB3 and ErbB4, mediate responses to EGF family members (5). EGF binding induces dimerization of the EGF receptor resulting in activation of the protein tyrosine kinase signaling pathway. These receptors undergo a complex pattern of ligand-induced homo- or hetero-dimerization to transduce EGF family signals (6, 7). EGF binds ErbB1 and depending on the context, induces the formation of homodimers or heterodimers containing ErbB2. Dimerization results in autophosphorylation of the receptor at specific tyrosine residues to create docking sites for a variety of signaling molecules (5, 8). Biological activities ascribed to EGF include epithelial development, angiogenesis, inhibition of gastric acid secretion, fibroblast proliferation, and colony formation of epidermal cells in culture.
Harris, R.C. et al. (2003) Exp. Cell Res. 284:2.
Carpenter, G. and Cohen, S. (1990) J. Biol. Chem. 265:7709.
Bell, G.I. et al. (1986) Nucl. Acids Res. 14:8427.
Carpenter, G. and Zendegui, J.G. (1986) Exp. Cell Res. 164:1.
Jorissen, R.N. et al. (2003) Exp. Cell Res. 284:31.
Gamett, D.C. et al. (1997) J. Biol. Chem. 272:12052.
Qian, X. et al. (1994) Proc. Natl. Acad. Sci. 91:1500.
Currently there are no citations for Recombinant Human EGF GMP Protein, CF.
The vial is supposed to contain lyophilized protein but it appears to be empty. Is there anything in it?
Pellets can be dislodged during shipping and become disbursed on the vial wall and in the cap. Centrifuge or tap the vial on the benchtop to return this material to the vial bottom. If this does not reveal a pellet, closely inspect the cone of the vial. Some pellets appear as only a tiny amount of material or as a transparent film due to the original buffer formulation. This is a normal appearance for many proteins. For example, if the product is originally lyophilized from a solvent such as acetonitrile or ethanol, and supplied carrier-free, you may not be able to detect the pellet with the naked eye. This does not mean the vial is empty. Reconstitute the vial as directed. After reconstitution, protein concentration can be tested with a spectrophotometer.
What is the recommended method for reconstitution of a lyophilized protein or antibody?
Unless more specific directions are on the Certificate of Analysis provided with the product, we suggest the following procedure to ensure optimal recovery: 1. Allow the vial and reconstitution buffer to equilibrate to room temperature. 2. Briefly centrifuge the vial to ensure that all lyophiliate is collected at the bottom of the vial. 3. Add the amount of buffer required to achieve the concentration recommended on the product insert. 4. Allow the vial to reconstitute for 15-30 minutes at room temperature with gentle agitation, like on a rocker platform or rotating by hand. Avoid vigorous shaking that can cause foaming and protein denaturation. 5. Aliquot into volumes greater than 20 μL and store as indicated on the product insert. If the vial exhibits flakes or particulates, mix the product for a couple of hours at room temperature and then at 4oC overnight. Contact Technical Service if product does not go into solution.