UA-Glo® Enhanced Bright Luciferase Assay System
UA-Glo® Enhanced Bright Luciferase Assay System
Product Details
Product Details
Product Specification
| Stability & Storage | Dry ice transport. Store protected from light at −20°C or lower; for long-term (>3 months) storage, −80°C is recommended. Refer to the reagent vial label for the expiration date. After first use, it is advisable to aliquot the reagent and store it protected from light at −20°C or lower. The reagent can be frozen and thawed no more than five times. Storing the reagent at 4°C for 24 hours or at room temperature (22°C) for 5 hours does not affect its function, with activity retained at >90%. |
Background
The UA-Glo® Enhanced High-Brightness Luciferase Assay Kit is used to quantitatively measure the expression level of the reporter gene luciferase in cells. Its homogeneous, ready-to-use formulation requires only a single addition of reagent for detection, minimizing experimental errors that can arise from multiple pipetting steps. This product offers high sensitivity and a signal half-life exceeding 0.5 hours, making it particularly well-suited for high-throughput assays of samples with low luciferase expression levels.
Components
UA-Glo® EnhancedThe components and specifications of the high-brightness luciferase assay kit are as follows. The number of reactions that can be detected in 96-/384-well plates is calculated based on adding 100 μL/20 μL of the assay reagent per well.
Specifications |
Number of reactions in a 96-well plate |
Number of reactions in a 384-well plate |
10 mL |
100 |
500 |
100 mL |
1,000 |
5,000 |
10 x 100 mL |
10,000 |
50,000 |
Protocol
UA-Glo® Enhanced High-Sensitivity Luciferase Assay Kit detects the activity of firefly luciferase reporter genes in cells through the luciferase reaction. The luciferase reaction is extremely sensitive, and parameters such as pH, temperature, and experimental system setup can significantly affect the reaction; therefore, direct comparison of absolute sample readings is generally not recommended under most experimental conditions. It is advised to set up the same reference control on each assay plate and to normalize the sample readings against both the reference control and the solvent carrier control before data processing and comparison.
1. According to experimental requirements, seed experimental cells expressing the firefly luciferase reporter gene at an appropriate density into 96-well or 384-well white transparent cell culture plates.
2. After performing cell treatments as needed, detect the luciferase expression levels in the cells.
3. Remove the UA-Glo®EnhancedHigh-sensitivity luciferase assay reagent. After the assay reagent has completely melted, equilibrate it to room temperature (22°C–25°C), then invert thoroughly to mix.
4. Remove the experimental cell plate and equilibrate it to room temperature. The luciferase reaction is sensitive to temperature changes. Both the assay reagent and the cell plate must be brought to room temperature, and the test should be conducted at a constant temperature (±1°C).
5. Add an equal volume of UA-Glo® high-sensitivity luciferase assay reagent to each well of the experimental cell plate as the culture medium in that well—for example, add 100 µL of the assay reagent to cells in a 96-well plate containing 100 µL of medium, or add 20 µL of the assay reagent to cells in a 384-well plate containing 20 µL of medium.
6. Shake the plate at medium speed for 1 minute, then incubate in the dark for 10 minutes to stabilize the luminescence signal.
7. Read the luminescence signal on a luminometer or multifunctional microplate reader, preferably within 30 minutes.
Guidelines
Without rigorous validation, it is not advisable to alter the volume of the test reagent.
For research use only.
