Key features and details
- Detection method: Luminescent
- Platform: Microplate reader
- Sample type: Adherent cells, Cell culture extracts, Cell culture media
Product nameLuciferase Reporter Assay Substrate Kit - Firefly
See all Luciferase Reporter kits
Sample typeCell culture extracts, Adherent cells, Cell culture media
Luciferase Reporter Assay Substrate Kit - Firefly (ab228530) provides a fast, simple, and homogeneous bioluminescence assay for the quantification of luciferase activity in live cells and cell extracts. This assay is based on firefly luciferase, a monomeric 61 kD enzyme that catalyses a two-step oxidation of luciferin, which yields light at 560 nm. The assay can be performed in a convenient 96-well and 384-well microtiter-plate format. The signal with a half-life of two to four hours provides a consistent signal across large batches of plates. The assay is compatible with the use of standard cell growth media. It has high sensitivity, and can be used for the assays that require low detection limit.
Common reporter genes include ß-galactosidase, ß-glucuronidase and luciferase. The advantages of a luciferase assay include: high sensitivity, absence of luciferase activity inside most of the cell types, wide dynamic range, speed and low cost. The most versatile and common reporter gene is the luciferase of the North American firefly Photinus pyralis. The protein requires no posttranslational modification for enzyme activity. It is not even toxic in high concentration (in vivo) and can be used in pro- and eukaryotic cells. The firefly luciferase catalyzes the bioluminescent oxidation of luciferin in the presence of ATP, magnesium and oxygen.
Storage instructionsStore at -20°C. Please refer to protocols.
Components 1 units Assay Buffer 1 x 10ml Luciferase Sensor 1 vial
Luciferase dose response was measured with Luciferase Reporter Assay Substrate Kit - Firefly (ab228530) in a white 96-well plate. The kit can detect as low as 0.1 pg/well luciferase with 20 minutes to 5 hours incubation without losing signal intensity. The integration time was 1 second. The half life is more than 4 hours.
Reaction Kinetics of CHO-V2R-Luc cells using Luciferase Reporter Assay Substrate Kit - Firefly (ab228530). CHO cells stably transfected with pCRE-luciferase gene and human Vasopressin receptor 2 (V2R) were plated into a white wall/clear bottom 384-well plate at 15,000 cells/well/25 µL. Cells then were treated with 100 nM of vasopressin in a 37 °C, 5% CO2 incubator for 4 hours. 25 µL of luciferase assay solution was added into the well. The kinetic data was taken every 30 minutes for up to 3 hours. The vasopressin induced luciferase signal is stable for more than 3 hours.
ab228530 has not yet been referenced specifically in any publications.