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Analysis Affinity of LSN2478185 and LY2495655 with Myostatin by KinExA (CAT#: STEM-MB-0027-CJ)

Introduction

Cancer cachexia is a multifactorial syndrome defined as persistent loss of skeletal muscle mass (with or without loss of fat mass) that cannot be fully reversed with regular nutritional support and leads to progressive sexual dysfunction. Myostatin has been identified as a negative regulator of skeletal muscle mass from the muscular hyperplasia phenotypes of naturally occurring subtypes and knockout mice. Reduction or overexpression of myostatin can lead to skeletal muscle hypertrophy and atrophy, respectively. Increased myostatin levels and pathway activation were found in preclinical models of cancer cachexia as well as in atrophic muscles of cancer patients. LSN2478185 is a myostatin neutralizing murine IgG1 monoclonal antibody and its derivative LY2495655, which is humanized and neutralizing monoclonal antibody against myostatin and significantly alleviates loss of skeletal muscle mass and function associated with tumor load.




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, Toxicology; 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 Proteins, DNA, Viruses, Serum, & More
• Equipment: Kinetic Exclusion Assay (KinExA)
• Optionals: CHO Cells
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