MBI Evolution LUCY Luciferase Assay Kit

• Luciferase Assay Kit is designed for cell biology to study gene expression and other cellular events related to gene expression.
• Gene expression is generally studied by linking a promoter sequence to an easily detectable "reporter" gene such as that encoding for firefly luciferase.
• Firefly luciferase catalyzes the oxidative carboxylation of luciferin, reacting with the highest efficiency of any known bioluminescence reaction.
• The assay is very sensitive because its light production has the highest quantum efficiency known for any chemiluminescent reaction, and no background  luminescence is found in the host cells or the assay chemistry.
• The assay is rapid, requiring only a few seconds per sample.

The Luciferase Assay is substantially improved over conventional assay methods in both sensitivity and simplicity. Light is produced by converting the chemical energy of luciferin oxidation through an electron transition, forming the product molecule oxyluciferin. Firefly luciferase is a 62kDa protein which is active as a monomer and does not require subsequent processing for its activity.

However, several factors may affect the sensitivity and success of the assay including pH, temperature, and substrate concentration. To ensure maximum sensitivity, the assay is performed in the presence of excess ATP, luciferin and Mg2+ in a buffer that will maintain a pH of 7.8. In the conventional assay for luciferase, a flash of light is generated that decays rapidly after the enzyme and substrates are combined.

Generally, 100-fold greater sensitivity can be achieved over the chloramphenicol acetyltransferase (CAT) assay. The Luciferase Assay System was developed for reporter quantitation in mammalian cells .

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