Description
Principle of the Assay: This assay employs a two-site sandwich ELISA to quantitative AGEs in Human serum, plasma, tissue homogenates, cell lysates. An antibody specific for AGEs has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and any AGEs present is bound by the immobilized antibody. After removing any unbound substances, a biotin - conjugated antibody specific for AGEs is added to the wells. After washing, Streptavidin conjugated Horseradish Peroxidase (HRP) is added to the wells. Following a wash to remove any unbound avidin-enzyme reagent, a substrate solution is added to the wells and color develops in proportion to the amount of AGEs bound in the initial step. The color development is stopped and the intensity of the color is measured.
Background: In human nutrition and biology, advanced glycation end products, known as AGEs, are substances that can be a factor in the development or worsening of many degenerative diseases, such asdiabetes, atherosclerosis, chronic renal failure, and Alzheimer's disease. These harmful compounds can affect nearly every type of cell and molecule in the body and are thought to be one factor in aging and in some age-related chronic diseases. They are also believed to play a causative role in the blood-vessel complications of diabetes mellitus. AGEs are seen as speeding up oxidative damage to cells and in altering their normal behavior. AGEs are formed both outside and inside the body. Specifically, they stem from glycation reaction, which refers to the addition of a carbohydrate to a protein without the involvement of anenzyme. Glucose can bind with proteins in a process called glycation, making cells stiffer, less pliable and more subject to damage and premature aging. Outside the body, AGEs can be formed by heating (for example, cooking). Intermediate products in the formation of an AGE are known as Amadori, Schiff base, and Maillard products, named after the researchers who first described them