Analysis Affinity of HGF and AMG 102 by Kinetic. (CAT#: STEM-MB-0005-CJ)

Introduction

Hepatocyte growth factor [HGF; also known as scatter factor (SF) ] is the only known ligand for c-Met, a receptor tyrosine kinase expressed in epithelial tissues that plays an essential role in the growth and maintenance of cells. Signaling through HGF/SF:c-Met mediates a number of normal cellular functions, including proliferation, survival, motility, migration, angiogenesis, and morphogenesis. Abnormal signaling via the HGF/SF:c-Met pathway has been associated with many types of cancers, with evidence for both paracrine and autocrine activation of c-Met by HGF; expression of these molecules in tumors and elevated concentrations of circulating HGF have been associated with an increased risk of tumor metastasis and poor prognosis.

AMG 102 is a fully human monoclonal antibody that selectively targets and neutralizes hepatocyte growth factor/scatter factor (HGF/SF). AMG 102 also inhibited cynomolgus monkey HGF-induced migration of human MDA-MB-435 cells but not rat HGF-induced migration of mouse 4T1 cells.

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Principle Applications Procedure Materials Notes Related Products

Principle

KinExA is a two-stage analysis system. In the first stage, a number of solutions are prepared, where one partner remains constant (constant binding partner, or CBP) and the other (titrant) is variable, usually serially diluted. As the titrant is added, the free CBP decreases and is analysed by a sophisticated and precise microfluorescence measurement device. The signal generated can be mathematically related to the affinity (KD) of the two molecules for each other, as well as the kinetic parameters of binding (kon) and dissociation (koff).

Applications

Oncology & Cancer, Metabolic; Immunology/Inflammation, Toxicology

Procedure

1. preparation of the functionalized beads which will capture the analyte for measuremen.
2. preparation of a series of solutions consisting of a constant initial concentration of one component of the binary reaction and serial dilutions of the other reactant. The component that is kept constant is the constant binding partner (CBP) , and is the one which will be analyzed.
3. each reaction mixture is sampled and the fluorescence of free CBP bound to the capture beads is obtained for subsequent numerical analysis.

Materials

• Sample Type: Antibodies, Cells, Small Molecules, Proteins, DNA, Lipids, Viruses, Serum, & More
• Equipment: Kinetic Exclusion Assay (KinExA)

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