Analysis Affinity of 39F7 to Human β-Klotho by KinExA (CAT#: STEM-MB-0047-CJ)

Introduction

Fibroblast growth factor 21 (FGF21) , an endocrine hormone in the FGF family, plays a key role in regulating metabolic homeostasis and has become a therapeutic target for metabolic diseases including type 2 diabetes. FGF21 acts through a receptor complex consisting of an FGF receptor (FGFR) and a co-receptor β-Klotho. 39F7, a high-affinity monoclonal antibody (mAb) against beta-Klotho, mimics the function of FGF21. In addition, the identification of 39F7 focused on β-Klotho's Trp-295 in a non-competitive manner with FGF21. In vitro characterization showed that 39F7 was specific for β-Klotho/FGFR1c activation. In addition, 39F7 can act by promoting receptor/co-receptor dimers.

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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

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, Serum, & More
• Equipment: Kinetic Exclusion Assay (KinExA)

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