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.

Are R&D Systems recombinant proteins and antibodies sterile?

Although the vials are bottled using aseptic techniques, heat-treated vials, and sterile stock solutions, they are not considered or guaranteed to be sterile. If sterile material is needed for an experiment, the material can be filtered through a 0.2 micron filter designed for use with biological fluids.

Recombinant Human EGF, Animal-Free Protein

Name: Recombinant Human EGF, Animal-Free Protein
Brand: R&D Systems
SKU: AFL236

R&D Systems, Inc. a Bio-Techne Brand

Catalog # AFL236

Citations (5)

Product Datasheet / COA / SDS

Catalog #

Availability

Size / Price

Qty

Setting the standard in quality research reagents

Key Product Details

Learn more about Animal-Free Recombinant Proteins

Source:	E. coli
Accession #:	P01133
Applications:	Bioactivity

View Full Technical Details for Recombinant Human EGF, Animal-Free Protein

Product Specifications for Recombinant Human EGF, Animal-Free Protein

Source	E. coli-derived human EGF protein Asn971-Arg1023, with an N-terminal Met Produced using non-animal reagents in an animal-free laboratory.
Purity	>97%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.
Endotoxin level	<0.10 EU per 1 μg of the protein by the LAL method.
N-terminal sequence Analysis	Met
Predicted Molecular Mass	6 kDa
Activity	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).

Formulation, Preparation and Storage

Carrier Free

What does CF mean?

CF stands for Carrier Free (CF). We typically add Bovine Serum Albumin (BSA) as a carrier protein to our recombinant proteins. 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 could interfere.

AFL236

Formulation	Lyophilized from a 0.2 μm filtered solution in PBS.
Reconstitution	Reconstitute at 200 μg/mL in sterile PBS.
Shipping	The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.
Stability & 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.

Reconstitution Calculator

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

Volume (to add to vial)

Mass (in vial)

Desired Reconstitution Concentration

Background: EGF

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.

Item	Value
References	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. Qian, X. et al. (1999) J. Biol. Chem. 274:574.
Long Name	Epidermal Growth Factor
Entrez Gene IDs	1950 (Human); 13645 (Mouse); 25313 (Rat)
Alternate Names	EGF, HOMG4, URG, Urogastrone, beta-urogastrone, epidermal growth factor, epidermal growth factor (beta-urogastrone), hEGF, pro-epidermal growth factor

Manufacturing Specifications

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

Citations for Recombinant Human EGF, Animal-Free Protein (5)

Showing 1 - 5 of 5 citations Showing All

Filter By:

Search Title:

Citations are publications that use Bio-Techne products. Selected citations for Recombinant Human EGF, Animal-Free Protein include:

Species: Human
Sample Types: Whole Tissue
Applications: Cell Culture

S Ding et al. (2022), Patient-derived micro-organospheres enable clinical precision oncology Cell Stem Cell, 0(0).
PMID: 35508177
Species: Human
Sample Types: Whole Cells
Applications: Bioassay

M Berlak et al. (2022), Mutations in ALK signaling pathways conferring resistance to ALK inhibitor treatment lead to collateral vulnerabilities in neuroblastoma cells Molecular Cancer, 21(1):126.
PMID: 35689207
Species: Human
Sample Types: Whole Cells
Applications: Cell Culture

MJ Baker et al. (2020), Evaluation of active Rac1 levels in cancer cells: a case of misleading conclusions from immunofluorescence analysis J. Biol. Chem., 0(0).
PMID: 32817335
Species: Human
Sample Types: Whole Cells
Applications: Bioassay

MJ Baker et al. (2020), Evaluation of active Rac1 levels in cancer cells: a case of misleading conclusions from immunofluorescence analysis J. Biol. Chem., 0(0).
PMID: 32817335
Species: Human
Sample Types: Whole Cells
Applications: Bioassay

YH Cui et al. (2018), FBXL14 abolishes breast cancer progression by targeting CDCP1 for proteasomal degradation Oncogene, 0(0).
PMID: 29973690

There are no citations that match your criteria.

Showing 1 - 5 of 5 citations Showing All

FAQs for Recombinant Human EGF, Animal-Free Protein

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 4^oC overnight. Contact Technical Service if product does not go into solution.
Are R&D Systems recombinant proteins and antibodies sterile?

Although the vials are bottled using aseptic techniques, heat-treated vials, and sterile stock solutions, they are not considered or guaranteed to be sterile. If sterile material is needed for an experiment, the material can be filtered through a 0.2 micron filter designed for use with biological fluids.

Recombinant Human EGF, Animal-Free Protein

R&D Systems, Inc. a Bio-Techne Brand

Key Product Details

Customers also Viewed

Customers also Purchased

Product Specifications for Recombinant Human EGF, Animal-Free Protein

Formulation, Preparation and Storage

Carrier Free

Reconstitution Calculator

Background: EGF

Manufacturing Specifications

Citations for Recombinant Human EGF, Animal-Free Protein (5)

FAQs for Recombinant Human EGF, Animal-Free Protein

Product Documents for Recombinant Human EGF, Animal-Free Protein

Product Datasheets

COA

Certificate of Analysis

SDS

Bio-Techne Brands

Stay Connected