Analysis Affinity of Anti-IL-8 mAbs by KinExA (CAT#: STEM-MB-0009-CJ)

Introduction

Interleukin 8 (IL-8) , like IL-6, is secreted by macrophages and a variety of cells that express Toll-like receptors in response to the stimulation of pathogens. IL-8's primary function is to recruit neutrophils and other target cells through chemotaxis to the infected site to eliminate pathogens. IL-8 causes increased intracellular Ca2+, release of reactive oxygen species, and other physiological changes required for migration and phagocytosis. IL-8 is also known to promote angiogenesis. Agents that block the binding of IL-8 to its receptor have been shown to block inflammation in animal models of disease. And IL-8 may prove efficacious in treating multiple human diseases.

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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;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 (Human and Mouse)
• Equipment: Kinetic Exclusion Assay (KinExA)

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