Analysis Immunoassay of 2,4-Dinitrophenol (DNP) by KinExA (CAT#: STEM-MB-0084-CJ)

Introduction

2, 4-dinitrophenol (DNP) is an organic compound that, when left in a dry state, is a high explosive with a risk of instantaneous explosion. Its use as a diet aid has had serious side effects, leading to many deaths. In living cells, DNP acts as a proton ion carrier and is commonly used in biochemical studies to help explore the bioenergetics of chemoosmosis and other membrane transport processes.

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

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