Recombinant Human EGF, Animal-Free Protein

• Reactivity: Hu
• Applications: Bioactivity
• Format: Carrier-Free

Recombinant Human EGF, Animal-Free Protein Summary

• Additional Information: Intended for preclinical researchers who may transition to GMP EGF for their clinical work
• Details of Functionality: 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).
• Source: E. coli-derived human EGF protein
  Asn971-Arg1023, with an N-terminal Met
  Produced using non-animal reagents in an animal-free laboratory.
• Accession #: P01133
• N-terminal Sequence: Met
• Protein/Peptide Type: Animal-Free Recombinant Proteins
• Gene: EGF
• Purity: >97%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.
• Purity Statement: Antigen Affinity-purified
• Endotoxin Note: <0.10 EU per 1 μg of the protein by the LAL method.

Applications/Dilutions

• Dilutions:
  • Bioactivity
• Theoretical MW: 6 kDa.
  Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.

Packaging, Storage & Formulations

• Storage: Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 3 months, -20 to -70 °C under sterile conditions after reconstitution.
• Buffer: Lyophilized from a 0.2 μm filtered solution in PBS.
• Purity: >97%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.
• Reconstitution Instructions: Reconstitute at 200 μg/mL in sterile PBS.

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for Recombinant Human EGF, Animal-Free Protein

• beta-urogastrone
• EGF
• epidermal growth factor (beta-urogastrone)
• epidermal growth factor
• hEGF
• HOMG4
• pro-epidermal growth factor
• URG
• Urogastrone

Background

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.

1. Harris, R.C. et al. (2003) Exp. Cell Res. 284:2.
2. Carpenter, G. and Cohen, S. (1990) J. Biol. Chem. 265:7709.
3. Bell, G.I. et al. (1986) Nucl. Acids Res. 14:8427.
4. Carpenter, G. and Zendegui, J.G. (1986) Exp. Cell Res. 164:1.
5. Jorissen, R.N. et al. (2003) Exp. Cell Res. 284:31.
6. Gamett, D.C. et al. (1997) J. Biol. Chem. 272:12052.
7. Qian, X. et al. (1994) Proc. Natl. Acad. Sci. 91:1500.
8. Qian, X. et al. (1999) J. Biol. Chem. 274:574.

Manufacturing Process

Animal-Free Manufacturing Conditions
Our dedicated controlled-access animal-free laboratories ensure that at no point in production are the products exposed to potential contamination by animal components or byproducts. Every stage of manufacturing is conducted in compliance with R&D Systems' stringent Standard Operating Procedures (SOPs). Production and purification procedures use equipment and media that are confirmed animal-free.

Production

• All molecular biology procedures use animal-free media and dedicated labware.
• Dedicated fermentors are utilized in committed animal-free areas.

Purification

• Protein purification columns are animal-free.
• Bulk proteins are filtered using animal-free filters.
• Purified proteins are stored in animal-free containers in a dedicated cold storage room.

Quality Assurance

• Low Endotoxin Level.
• No impairment of biological activity.
• High quality product obtained under stringent conditions.
• For ex vivo research or bioproduction, additional documentation can be provided.

Please read our complete Animal-Free Statement

 

Publications for EGF (AFL236)(4)

Publications using AFL236

• M Berlak, E Tucker, M Dorel, A Winkler, A McGearey, E Rodriguez-, BM da Costa, K Barker, E Fyle, E Calton, S Eising, K Ober, D Hughes, E Koutrouman, P Carter, R Stankunait, P Proszek, N Jain, C Rosswog, H Dorado-Gar, JJ Molenaar, M Hubank, G Barone, J Anderson, P Lang, HE Deubzer, A Künkele, M Fischer, A Eggert, C Kloft, AG Henssen, M Boettcher, F Hertwig, N Blüthgen, L Chesler, JH Schulte Mutations in ALK signaling pathways conferring resistance to ALK inhibitor treatment lead to collateral vulnerabilities in neuroblastoma cells Molecular Cancer, 2022-06-10;21(1):126. 2022-06-10 [PMID: 35689207] (Bioassay, Human)
• S Ding, C Hsu, Z Wang, NR Natesh, R Millen, M Negrete, N Giroux, GO Rivera, A Dohlman, S Bose, T Rotstein, K Spiller, A Yeung, Z Sun, C Jiang, R Xi, B Wilkin, PM Randon, I Williamson, DA Nelson, D Delubac, S Oh, G Rupprecht, J Isaacs, J Jia, C Chen, JP Shen, S Kopetz, S McCall, A Smith, N Gjorevski, AC Walz, S Antonia, E Marrer-Ber, H Clevers, D Hsu, X Shen Patient-derived micro-organospheres enable clinical precision oncology Cell Stem Cell, 2022-05-03;0(0):. 2022-05-03 [PMID: 35508177] (Cell Culture, Human)
• MJ Baker, M Cooke, G Kreider-Le, R Garcia-Mat, PA Janmey, MG Kazanietz Evaluation of active Rac1 levels in cancer cells: a case of misleading conclusions from immunofluorescence analysis J. Biol. Chem., 2020-08-14;0(0):. 2020-08-14 [PMID: 32817335] (Bioassay, Cell Culture, Human)
• YH Cui, H Kim, M Lee, JM Yi, RK Kim, N Uddin, KC Yoo, JH Kang, MY Choi, HJ Cha, OS Kwon, IH Bae, MJ Kim, N Kaushik, SJ Lee FBXL14 abolishes breast cancer progression by targeting CDCP1 for proteasomal degradation Oncogene, 2018-07-04;0(0):. 2018-07-04 [PMID: 29973690] (Bioassay, Human)

Bioinformatics

• Gene Symbol: EGF
• Uniprot:
  • P01133()