Analysis Affinity of Admium-EDTA Complexes Antibodies by KinExA (CAT#: STEM-MB-0038-CJ)

Introduction

Disodium EDTA is a chelating agent used in calcium studies, and EDTA is thought to be aggressive against calcium homeostasis mechanisms. The sodium ions of the chelate are replaced by calcium, which can form soluble, non-toxic, and physiologically unavailable complexes that are excreted in the urine. When the EDTA combines with available calcium in the blood, the animal is forced into a negative calcium equilibrium. In this case, the calcium homeostasis mechanism may respond to skeletal, gastrointestinal, and renal effects via parathyroid hormone and 1,25 (OH) 2 vit D3.

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

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