Analysis Affinity of Met-pep1 to Met by KinExA (CAT#: STEM-MB-0011-CJ)

Introduction

C-Met and its gene product Met have been extensively studied in terms of their relevance to cancer biology. Activation of Met via autocrine, paracrine, or mutational mechanisms can lead to tumorigenesis and metastasis. Numerous studies have linked inappropriate expression of this ligand-receptor pair to most types of human solid tumors, including those of brain, breast, ovary, thyroid, pancreas, stomach, prostate, and nasopharyngeal carcinoma.Clinical applications of a Met binding peptide (MET-PEP) as a diagnostic reagent and therapeutic vector can be sought.

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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; Immunology/Inflammation; Pharmacology

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, Lipids, Serum
• Equipment: Kinetic Exclusion Assay (KinExA)

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