Figure 1. hEGF levels in normal patient serum, plasma and urine samples were quantified using SignalKine™ Human EGF Sandwich ELISA Kit.
Typical Data. Represented below are hEGF Standard Curves, one diluted in SignalKine™ Sample Diluent S02, a second diluted in cell culture media (RPMI + 10% FBS). Although these are typical of the standard curves that will be generated using this kit, a new set of standards should be run for each new experiment.
SignalKine™ Human EGF Sandwich ELISA Kit from Cell Signaling Technology (CST) is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects human EGF (hEGF) in multiple matrices. Unknown samples being tested for hEGF and hEGF Standards are added to low volume microwells, where the hEGF is captured by the coated hEGF Rabbit mAb. Following a washing step, a biotinylated hEGF Rabbit mAb is added to detect the captured hEGF. HRP-linked Streptavidin is then used for detection of the biotinylated hEGF Detection Rabbit mAb. HRP substrate, TMB, is added for color development. The magnitude of absorbance for this developed color is proportional to the quantity of EGF in the sample.
SignalKine™ Human EGF Sandwich ELISA Kit detects hEGF in multiple matrices that be quantified by generating a standard curve with the recombinant hEGF protein standard provided. The hEGF standard range is from 3.9 to 250 pg/ml. Samples containing higher levels of hEGF can be diluted to fit into the standard range.
Epidermal growth factor (EGF) is a small polypeptide hormone that has mitogenic properties, in vivo and in vitro, and affects the growth and/or differentiation of many cell types. EGF elicits biologic responses by binding to its cell surface receptor, which is a transmembrane glycoprotein containing a cytoplasmic protein tyrosine kinase (1,2). The binding of EGF to the EGF receptor promotes dimerization of the receptor, autophosphorylation, and activation of downstream signaling components (3). The integrated biological responses to EGF signaling are pleiotropic including mitogenesis, apoptosis, enhanced cell motility, protein secretion, differentiation, and dedifferentiation. In addition to being implicated in organ morphogenesis, maintenance, and repair, research studies have correlated upregulated EGF receptor signaling with progression to invasion and metastisis in a wide variety of tumors (4-6). Thus, investigators have identified EGF receptor and its downstream signaling molecules as potential targets for therapeutic interventions in wound repair and cancer (4-6).
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