UA-Glo® Enhanced Bio-luc Luciferase Assay System
UA-Glo® Enhanced Bio-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, we recommend −80°C. Refer to the reagent vial label for the expiration date. After first use, it is recommended 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. For short-term continuous use, the reagent may be stored at 4°C for no more than five days. |
Background
The UA-Glo® Enhanced Bio-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® Bio-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® EnhancedBio-luciferase fluorescence assay kit specifications 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 |
500 |
100 mL |
1,000 |
5,000 |
Protocol
UA-Glo® EnhancedBThe Bio-luciferase assay kit detects the activity of firefly luciferase reporter genes in cells through a luciferase reaction. The luciferase reaction is extremely sensitive, and parameters such as pH, temperature, and experimental system setup can significantly affect the reaction. Under most experimental conditions, it is not recommended to directly compare absolute detection values of samples. It is suggested to set up the same reference control on each experimental plate, and to normalize the detection values of experimental samples using the reference control and 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 treatment according to experimental needs, proceed with the luciferase reporter gene assay.
3. Remove the UA-Glo®EnhancedBio-luc luciferase assay reagent. After the assay 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 should be conducted at a constant temperature (±1°C).
4. Remove the experimental cell plate and equilibrate it 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 place it in the dark and incubate 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.
