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Analysis Immunoassay of Polychlorinated Biphenyls (PCBs) by KinExA (CAT#: STEM-MB-0080-CJ)

Introduction

Polychlorinated biphenyls (PCBs) are a group of manmade chemicals. They are oily liquids or solids, clear to yellow in color, with no smell or taste. PCBs are very stable mixtures that are resistant to extreme temperature and pressure. PCBs were used widely in electrical equipment like capacitors and transformers. They also were used in hydraulic fluids, heat transfer fluids, lubricants, and plasticizers.

In people, PCBs can affect the skin and may cause chloracne--small, pale, yellow skin lesions that may last from weeks to years. PCBs also can cause short-term changes in the activity of the liver, but without any noticeable symptoms. These liver changes are similar to those resulting from the consumption of alcoholic beverages or smoking cigarettes. Animal studies also have suggested that PCBs can affect the immune, endocrine and reproductive systems, but these effects are uncertain in humans. Large amounts of PCBs given to laboratory animals over a short time can cause cancer. Studies of human workers exposed to high levels of PCBs for long periods have not consistently shown that PCBs cause cancer in humans.Birth defects have been also linked to mothers who have been exposed to PCBs.




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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