One step firefly luciferase reporter gene detection kit

Reported gene testing is an important tool in modern molecular biology research to analyze the potential interactions between cis elements (such as promoters, enhancers, and silencers) and trans acting factors in the lateral regions of structural genes.
Firefly luciferase is widely used in gene regulation and drug screening. Firefly luciferase is a protein with a molecular weight of approximately 61 kD. In the presence of ATP, magnesium ions, and oxygen, it can catalyze the production of oxyluciferin from luciferin. During the oxidation of luciferin, a light signal is generated, as shown in Figure 1.
The light signal of this reagent kit is a momentary light, which needs to be detected immediately after adding the working solution. The half-life of the optical signal is about 5 minutes.

Product components:

1. Working fluid configuration
(1) Restore all components to room temperature.
(2) Dilute component B (storage solution) thoroughly with component A to prepare a 0.2 mg/mL firefly luciferase working solution, vortex and shake to ensure thorough mixing.
Note: Firefly luciferase working solution cannot be repeatedly frozen and thawed. If the amount used in a single experiment is small, it is recommended to divide it according to the amount used in a single experiment. At room temperature, after 3 hours of preparation of the working fluid, the activity decreased by about 10%, and after 5 hours, the activity decreased by about 25%.
2. Chemiluminescence value detection
(1) Take out the cell culture plate from the incubator and incubate at room temperature for 20 minutes to restore it to room temperature (22-25 ° C).
(2) Add an equal volume of firefly luciferase working solution to the culture plate and mix well.
(3) Incubate at room temperature for 5 minutes.
Note: The incubation time can be adjusted appropriately according to the cell type and number.
(4) Read the values using a multifunctional enzyme-linked immunosorbent assay (ELISA) reader or chemiluminescence analyzer (instrument parameters: measurement time of 10 seconds, measurement interval of 2 seconds).

-Store at 80 ℃. B component is recommended to be pre prepared with sterile water as a 2 mg/mL storage solution. A component and B component prepared as a storage solution should be prepared according to the experiment
Require small batch packaging. It is recommended to prepare and use the testing fluid immediately to avoid repeated freezing and thawing.

1. Before use, please centrifuge the product instantly to the bottom of the tube before conducting subsequent experiments.
2. The strongest wavelength of bioluminescence catalyzed by firefly luciferase is 560 nm.
3. To prevent interference between holes, it is recommended to use white opaque orifice plates.

Firefly luciferase reporter gene detection kit, one-step firefly luciferase reporter gene detection kit