Luciferin and luciferase: a brilliant performance!
D-Luciferin and the enzyme luciferase are a well-established team in reporter gene assays and a useful tool in visualizing cellular processes.
D-Luciferin ((4S)-4,5-Dihydro-2-(6-hydroxy-2-benzothiazolyl)-4-thiazolecarboxylic acid) and its salts are chemiluminescent substrates of the enzyme luciferase. They are also known as firefly luciferin or beetle luciferin.
Catalyzed by the enzyme luciferase, the membrane-permeable and water-soluble D-luciferin is first converted to luciferin adenylate and finally degraded to oxyluciferin under light emission. The reaction requires ATP and oxygen.
The enzyme luciferase is a well-established reporter in plants, bacteria, and mammalian cells. Luciferase reporter assays are a classical method to study regulatory DNA regions: When the luciferase gene is inserted downstream of a promoter into a coding DNA segment, the intensity of the bioluminescence produced allows conclusions regarding the activity of the promoter/gene.
Living cells, tissues, animal models
D-Luciferin is used for bioluminescent imaging of living cells, tissues and animal models. The range of applications of the luciferase/D-luciferin system is very similar to that of fluorescent proteins. However, the luciferase/D-luciferin system has the advantage of operating with virtually no background – emitted light is detected in a previously light-free space. In addition, compared to fluorescence-based reporter assays, luciferase-based assays have significantly higher sensitivity and a higher linear range of analysis.
The main applications of D-luciferin (in combination with luciferase-expressing cells) are:
- to study intracellular signaling activity in vitro and in vivo
- to measure transfection/transduction efficacy
- to evaluate and measure of gene expression
- to study gene regulation
- for promotor analysis
- for compound screenings
- to study protein-protein interactions (Luciferase Complementation Assay)
- in ATP assaya
- in immunoassays
- for detection of very small amounts of ATP or oxygen
For more information on our D-Luciferins and related articles, please refer to the corresponding data sheets in our webshop!