UA-Glo® Renilla Luciferase Assay System
UA-Glo® Renilla Luciferase Assay System
Product Details
Product Details
Product Specification
| Synonyms | Sea Pansy Luciferase Assay Kit |
| Stability & Storage | Dry ice transportation. The lysis buffer should be stored in a dark place at -20℃ or below, and the sea kidney luciferase substrate should be stored at -80℃. The validity period is indicated on the bottle label. After the first use, it is recommended to aliquot the lysis buffer and the sea kidney luciferase substrate as needed and store them separately in the dark at -20℃ and -80℃, respectively. |
Background
The UA-Glo® Renilla Luciferase Assay Kit is used for quantitative detection of Renilla luciferase expressed in cells. The kit is a homogeneous reagent and can be tested with just one sample addition, reducing the experimental errors that may result from multiple sample additions. This reagent has the characteristics of high signal-to-noise ratio, good repeatability, and excellent stability. Its stable luminescent signal enables this product to be used for high-throughput sample detection.
Components
Renilla Luciferase Assay Buffer is packaged in 10 ml or 100 ml amber bottles, and the substrate is aliquoted into 2 ml tubes. Specifications are as follows:
| Buffer | Substrate | Detectable 96-well plate wells | Detectable 384-well plate wells |
| 10 ml | 100 μl | 100 wells | 500 wells |
| 100 ml | 1ml | 1,000 wells | 5,000 wells |
| 10x100 ml | 10x1ml | 1,000 wells | 50,000 wells |
Protocol
UA-Glo® Renilla Luciferase Assay Kit detects the activity of the Renilla luciferase reporter gene in cells through a luciferase reaction. The luciferase reaction is highly sensitive, and parameters such as pH, temperature, and experimental system settings have a significant impact on the reaction. Under most experimental conditions, direct comparison of absolute detection values between samples is not recommended. It is advised to set up the same reference control on each assay plate, and normalize the detection values of experimental samples against those of the reference control and vehicle control before data processing and comparison.
1) Seed the experimental cells expressing the Renilla luciferase reporter gene at an appropriate density in a white, transparent-bottom 96-well or 384-well cell culture plate according to experimental requirements. For example, seed 100 μL of cells per well in a 96-well plate.
2) After treating the cells as required by the experiment, proceed with the Renilla luciferase assay.
3) Remove the cell culture plate to be tested and equilibrate to room temperature (22°C–25°C). The luciferase reaction is sensitive to temperature changes. Both the assay reagent and the cell plate must be equilibrated to room temperature, and the temperature should be kept constant (±1°C) during the assay.
4) Take out the lysis buffer. After complete thawing, mix thoroughly by inverting, take the required amount, and equilibrate to room temperature.
5) Take out the Renilla luciferase substrate and add it to the lysis buffer at a 100× dilution to prepare the Renilla luciferase assay reagent.
6) Add an equal volume of the Renilla luciferase assay reagent to the culture medium in each well of the cell plate. For example, add 100 μL of assay reagent to wells of a 96-well plate containing 100 μL of cells, or 20 μL of assay reagent to wells of a 384-well plate containing 20 μL of cells.
7) Shake the plate at medium speed for 1 minute, then incubate in the dark for 10 minutes.
8) Measure the luminescence signal using a luminometer or a multimode plate reader. It is recommended to read the plate within 60 minutes.
Guidelines
1) It is not recommended to mix samples from different batches.
2) Without rigorous verification, it is not advisable to change the dosage of the detection reagents.
3) This is for research purposes only.
