The proliferation of NIH/3T3 cells treated with increasing concentrations of mEGF was assessed. After 24 hr treatment, cells were labeled with BrdU for 4 hr. BrdU incorporation was determined using the BrdU Cell Proliferation Assay Kit #6813 and the OD450 was determined.
The purity of recombinant mEGF was determined by SDS-PAGE of 6 µg reduced (+) and non-reduced (-) recombinant mEGF and staining overnight with Coomassie Blue.
Western blot analysis of extracts from NIH/3T3 cells, untreated or treated with mEGF for 10 minutes, using Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) (D13.14.4E) XP® Rabbit mAb #4370 (upper) or p44/42 MAPK (Erk1/2) (137F5) Rabbit mAb #4695 (lower).
With carrier: Lyophilized from a 0.22 μm filtered solution of 20 mM citrate, pH 3.0 containing 100 mM NaCl and 20 μg BSA per 1 μg mEGF. Carrier free: Lyophilized from a 0.22 μm filtered solution of 20 mM citrate, pH 3.0 containing 100 mM NaCl.
Stable in lyophilized state at -20°C for 1 year after receipt. Sterile stock solutions reconstituted with carrier protein are stable at 4°C for 2 months and at -20°C for 6 months. Avoid repeated freeze-thaw cycles.Maintain sterility. Storage at -20°C should be in a manual defrost freezer.
>98% as determined by SDS-PAGE of 6 μg reduced (+) and non-reduced (-) recombinant mEGF. All lots are greater than 98% pure.
Less than 0.01 ng endotoxin/1 μg mEGF.
The bioactivity of recombinant mEGF was determined in an NIH/3T3 cell proliferation assay. The ED50 of each lot is between 10-250 pg/ml.
Recombinant mEGF has a Met on the amino terminus and has a calculated MW of 6045. DTT-reduced and non-reduced protein migrate as 6 kDa polypeptides. The expected amino-terminal MNSYP of recombinant mEGF was verified by amino acid sequencing.
Recombinant mouse EGF (mEGF) Asn977-Arg1029 (Accession #NP_034243) was produced in E. coli at Cell Signaling Technology.
EGF is produced by epithelial cells, fibroblasts, and many other cell types (1,2). Low molecular weight soluble EGF is generated through proteolysis of a larger ~130,000 molecular weight transmembrane precursor (1,2). Both soluble and membrane forms of EGF are active (2). EGF induces proliferation, differentiation, and survival of many cell types including tumor-derived cells (1-3). There are multiple members of the EGF family and multiple members of the ErbB/HER EGF receptor family. EGF binds to ErbB1/HER1 and induces homodimerization or induces heterodimerization with other ErbB/HER members (1). Binding of EGF signals through the MAPK, PI3K/Akt, and Stat5 pathways (1). EGF, EGF family members, EGF receptors, and their signaling pathways are involved in many cancers and are targets for therapeutic intervention (1, 2).
Explore pathways + proteins related to this product.
Except as otherwise expressly agreed in a writing signed by a legally authorized representative of CST, the following terms apply to Products provided by CST, its affiliates or its distributors. Any Customer's terms and conditions that are in addition to, or different from, those contained herein, unless separately accepted in writing by a legally authorized representative of CST, are rejected and are of no force or effect.
Products are labeled with For Research Use Only or a similar labeling statement and have not been approved, cleared, or licensed by the FDA or other regulatory foreign or domestic entity, for any purpose. Customer shall not use any Product for any diagnostic or therapeutic purpose, or otherwise in any manner that conflicts with its labeling statement. Products sold or licensed by CST are provided for Customer as the end-user and solely for research and development uses. Any use of Product for diagnostic, prophylactic or therapeutic purposes, or any purchase of Product for resale (alone or as a component) or other commercial purpose, requires a separate license from CST. Customer shall (a) not sell, license, loan, donate or otherwise transfer or make available any Product to any third party, whether alone or in combination with other materials, or use the Products to manufacture any commercial products, (b) not copy, modify, reverse engineer, decompile, disassemble or otherwise attempt to discover the underlying structure or technology of the Products, or use the Products for the purpose of developing any products or services that would compete with CST's products or services, (c) not alter or remove from the Products any trademarks, trade names, logos, patent or copyright notices or markings, (d) use the Products solely in accordance with CST's Product Terms of Sale and any applicable documentation, and (e) comply with any license, terms of service or similar agreement with respect to any third party products or services used by Customer in connection with the Products.