With carrier: Add sterile 20 mM citrate, pH 3.0 to a final mEGF concentration of greater than 50 μg/ml. Solubilize for 30 minutes at room temperature with occasional gentle vortexing.
Carrier free: Add sterile 20 mM citrate, pH 3.0 or 20 mM citrate, pH 3.0 containing protein to minimize absorption of mEGF to surfaces. Solubilize for 30 minutes at room temperature with occasional gentle vortexing. Stock mEGF should be greater than 50 μg/ml.
|>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).
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