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Analysis Affinity of MMAb3 to NHP IL-13 and Human IL-13 by KinExA (CAT#: STEM-MB-0018-CJ)

Introduction

Interleukin 13 (IL-13) is a cytokine secreted from type 2 T helper (Th2) cells. IL-13 has overlapping functions with interleukin 4 (IL-4) , including the induction of immunoglobulin E (IgE) secretion from B cells, and the inhibition of interleukin 1 beta (IL-1Ž²) , tumor necrosis factor alpha (TNF-Ž±) , interleukin 8 (IL-8) , and interleukin 6 (IL-6) inflammatory cytokine expression. IL-13 also regulates immune cell.Nonhistone proteins (NHP) are associated with the scaffolding of DNA structures. They play an important role in regulating processes such as replication, transcription, nucleosome remodelling, etc。




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: Serum, Plasma, Urine
• Equipment: Kinetic Exclusion Assay (KinExA)
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