The MaxDiscovery™ Luciferase ELISA Kit is an enzyme immunoassay that analyzes the quantity of Luciferase in cells, serum and tissues. Luciferase is a generic name for enzymes, commonly found in nature, that produce light. The most commonly used Luciferase is produced by, Photinus pyralis, the American firefly, which can catalyze a chemiluminescent oxidation–reduction reaction. Although many kits are available to detect the enzymatic activity of this protein using light emission, these methods have several drawbacks. For example, these assays have rapid decay kinetics and require the enzymatic activity of the Luciferase to be retained. Since enzyme activity can be influenced by many factors including handling, storage and buffer conditions, these assays are more variable and more difficult to reproduce than ELISA based assays. In addition, enzyme based activity assays do not easily enable absolute quantification as enzyme activity is relative to a standard that does not contain equal representative activity with respect to the quantity of luciferase. In response to these issues, Bioo Scientific has developed a highly sensitive ELISA assay which allows the researcher to completely measure the amount of firefly Luciferase protein from a number of sources. Like most ELISA assays, the MaxDiscovery™ Luciferase ELISA kit relies on a Horseradish Peroxidase (HRP) conjugated Luciferase antibody and the TMB (3,3´,5,5´-tetramethylbenzidine) substrate. TMB is a chromogen that yields a blue color when oxidized with hydrogen peroxide (catalyzed by HRP) and has major absorbances at 370 nm and 652 nm. The color then changes to yellow with the addition of acid with maximum absorbance at 450 nm. The relative amount of Luciferase protein in the cells will be directly proportional to the amount of signal that is obtained at 450 nm.
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