Analysis Affinity of Human Anti-human Interleukin 6 (IL-6) Monoclonal Antibodies to IL-6 by Kinetic (CAT#: STEM-MB-0002-CJ)

Introduction

Interleukin-6 (IL-6) , a cytokine associated with many inflammatory diseases and many cancers, has a variety of beneficial effects when released as myocytokine by contracting muscles. In addition, IL-6 has an effect on drug metabolizing enzymes in human hepatocyte culture, and inhibits the activity of CYP1A2 and CYP3A4 enzyme markers in freshly isolated and cryopreserved hepatocytes. In addition, IL-6 inhibited the induction of omeprazole on CYP1A2 enzyme activity and rifampicin on CYP3A4 enzyme activity, but only at superphysiological IL-6 concentrations. There was no significant difference in the ability of glycosylated and non-glycosylated IL-6 to inhibit CYP1A2 and CYP3A4 enzyme activity. Anti-IL-6 monoclonal antibodies can eliminate or partially block the IL-6-mediated inhibition of CYP1A2 and CYP3A4 enzyme activity, respectively.

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

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