Signal stability test of UA-Glo Stable Luciferase Assay Kit: HEK293 cells were seeded into 96-well cell culture plates; after 5 hours, luciferase expression plasmids were transfected into the cells; 48 hours post-transfection, the cells were taken out and equilibrated at room temperature for 10 minutes, then UA-Glo Stable and High Luminance Luciferase Assay Reagent (equilibrated to room temperature) was added, mixed well, and the fluorescence values were continuously read at different time points.
UA-Glo® Steady Luciferase Assay System
UA-Glo® Steady Luciferase Assay System
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$38 USD
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Product Details
Product Details
Product Specification
| Synonyms | Stable Luciferase Assay Kit |
| Stability & Storage |
Reagents stored at 20°C that cannot be used up all at once after opening the bottle are recommended to be aliquoted and frozen at 20°C. It is recommended that freeze-thaw cycles do not exceed 3 times, and the time left at room temperature each time should not exceed 1 hour. |
Background
The UA-Glo Stable Luciferase Assay Kit is used for the quantitative detection of the content of stably expressed luciferase in cells. This reagent features high signal-to-noise ratio, good repeatability, and excellent stability. There is no need to remove the culture medium; the detection reagent can be directly added to the culture plate for detection, thereby further reducing operational errors. Its stable signal makes this product particularly suitable for high-throughput sample detection. The stable luciferase assay kit (Steady glo) has good reaction stability of the detection signal, with a half-life of more than 2 hours, which can meet the requirements of high-stability detection.
Components
ATP, luciferin, and buffer solution are mixed and then filled into 10 ml or 100 ml brown bottles. The specifications are as follows:
|
Product specifications |
Can detect the number of wells in a 96-well plate |
Can detect the number of wells in a 384-well plate |
|
10 ml |
100 |
500 |
|
100 ml |
1,000 |
5,000 |
|
10 x 100 ml |
10,000 |
50,000 |
Protocol
1. Before the experiment, equilibrate the UA-Glo stable luciferase assay reagent to room temperature and mix gently by shaking.
2. Equilibrate the cell plate to be tested (white opaque-bottom or black opaque-bottom plate) at room temperature for 10 minutes.
3. Add an equal volume of the assay reagent to that of the cell culture medium into each culture reaction well.
4. Shake on a plate shaker for 1 minute, then incubate the sample plate at room temperature in the dark for 5-8 minutes.
5. Read and record the fluorescence signal on a luminescence microplate reader.
Picture
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Bioactivity
*HEK293 cells were seeded into 6-well cell culture plates and transfected with luciferase expression plasmids; 48 hours post-transfection, the cells were digested with enzymes, counted, and then aliquoted into 96-well assay plates with the number of cells per well as shown in the table. Subsequently, UA-Glo Stable Assay Reagent (equilibrated to room temperature) was added for detection, with untransfected cells as controls. The fluorescence values were continuously read at different time points, and the reading ratios were calculated. The comparative reagent is a similar product from a well-known foreign brand.
Cell reporter gene assay using UA-Glo Stable Luciferase Assay Kit. Cells transfected with reporter gene vectors were plated into 384-well plates at a cell density of 5×10⁵ cells/mL. Compounds at different concentrations were added and the cells were cultured for 24 hours. Then, luciferase activity was detected using stable luciferase assay kits, including products from the international brand Px Company and UA-Steady products.
Compound A and Compound B represent compounds with strong activity and weak activity, respectively. The results showed that the differences in IC50 values between the two products were all within 2-fold.
