UA-Glo® Enhanced One-luc Luciferase Assay System
UA-Glo® Enhanced One-luc 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 may be frozen and thawed no more than five times. For short-term continuous use, the reagent can be stored at 4°C for no more than five days. |
Background
The UA-Glo® Enhanced One-luciferase Assay Kit is used to quantitatively measure the expression level of the reporter gene luciferase in cells. This kit features a homogeneous assay format, requiring only a single addition of reagents for detection, thereby minimizing experimental errors that may arise from multiple pipetting steps. The kit boasts a high signal-to-noise ratio, excellent reproducibility, and superior stability. Its stable glow-type luminescence makes it particularly well suited for high-throughput compound screening. The UA-Glo® One-luciferase assay reagent exhibits enhanced chemical stability; when stored at 4°C for five days, the reduction in signal intensity remains below 15%, making it convenient for customers who need to repeatedly use the kit within a short period.
Components
UA-Glo® EnhancedThe specifications for the One-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 wells in a 96-well plate |
Number of wells in a 384-well plate |
10 mL |
100 |
5,00 |
100 mL |
1,000 |
50,00 |
10 × 100 mL |
10,000 |
50,000 |
Protocol
UA-Glo® Enhanced One-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, under most experimental conditions, it is not recommended to directly compare absolute detection values of samples. It is advised to set the same reference control on each assay plate and to normalize the detection values of experimental samples using 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, proceed with the luciferase reporter gene assay.
3. Remove the UA-Glo®EnhancedOne-luciferase assay reagent. After the reagent has completely melted, equilibrate it to room temperature (22°C–25°C) and mix thoroughly by inverting. The luciferase reaction is sensitive to temperature changes. Both the assay reagent and the experimental cell plate should be brought to room temperature, and the test temperature should remain constant (±1°C).
4. Remove the experimental cell plate and allow it to equilibrate to room temperature.
5. Add an equal volume of assay reagent to each well of the experimental cell plate as the culture medium in that well; for example, add 100 µL of assay reagent to cells in a 96-well plate containing 100 µL of medium, and add 20 µL of 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 plate reader, and it is recommended to complete the reading within 1 hour.
Guidelines
Without rigorous validation, it is not advisable to alter the volume of the test reagent.
For research use only.
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Bioactivity
