Analysis Immunoassay of Chelator-Linked Antibodies by KinExA (CAT#: STEM-MB-0083-CJ)

Introduction

Monoclonal antibodies directed toward metal chelating agents or metal-chelate complexes have many potential uses both in medicine and in environmental analysis. In medicine, chelator-linked antibodies have been used in vivo to transport and deliver radioisotopes to specific target sites, such as tumors. The availability of monoclonal antibodies to the chelating agent has permitted researchers to investigate the biodistribution of these “magic bullets” in cancer chemotherapy. In environmental analysis, the availability of monoclonal antibodies that can distinguish among different metals provides a basis for rapid, sensitive immunoassays that can be used on-site to assess heavy metal contamination.

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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, Lipids, Serum (Human and mouse)
• Equipment: Kinetic Exclusion Assay (KinExA)

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