Product Details
Product Details
Product Specification
| Synonyms | Renilla luciferase detection kit |
| Stability & Storage |
Store the Renilla luciferase assay kit and aliquoted substrates at -20°C. The buffer can be stored at 4°C. |
Background
The UA-Glo Renilla Luciferase Assay Kit is used for the quantitative detection of Renilla luciferase content in cells or other samples. This reagent features high signal-to-noise ratio, good repeatability, and good stability. The signal half-life exceeds 2 hours. The simple formula reduces the experimental steps of lysing first and then detecting, thereby further reducing errors caused by frequent sample addition, and its stable glow signal makes this product particularly suitable for high-throughput sample detection.
Components
Renilla luciferase buffer is filled in 10 ml or 100 ml brown bottles, and the substrate is dispensed into 2 ml tubes. The specifications are as follows:
|
buffer solution |
substrate |
Detectable number of wells in 96-well plate |
Detectable number of wells in 384-well plates |
|
10 ml |
100 ml |
200 wells |
1,000 wells |
|
100 ml |
1ml |
2,000 wells |
10,000 wells |
|
10x100 ml |
10x1ml |
2,000 wells |
100,000 wells |
Protocol
1. Plate the cells to be tested at an appropriate density in a 96-well or 384-well cell culture plate, and transfect with appropriate plasmids or stable cell lines carrying the relevant Renilla luciferase expression vector. It is recommended to use a white plate.
2. Perform relevant treatments on the cells according to project requirements and continue culturing for an appropriate period of time.
3. Take out the detection buffer and substrate, and equilibrate at room temperature for 20 minutes.
4. Take out the cell culture plate to be tested and equilibrate at room temperature for 20 minutes.
5. Prepare the detection reagent. The substrate is a 100x concentrate, and it should be prepared into a 1x working solution with buffer before use. Note to avoid light.
6. Add 50 µl of detection reagent to 100 µl of cells in a 96-well plate, or 10 µl of reagent to 20 µl of cells in a 384-well plate, gently shake for 30 seconds, and place in the dark to continue lysis for 3 minutes.
7. Read the fluorescence signal on a luminescence microplate reader.
8. It is recommended to store unused reagents at -20℃.
Guidelines
1. Temperature: Unless otherwise specified, luciferase detection is carried out at room temperature (22-25°C). Temperature has a significant impact on enzyme reactions.
2. Reagent dosage: It is not recommended to arbitrarily change the dosage of reaction reagents without strict verification.
3. Plate reading time: After adding the reagents, shake for 30 seconds, place in the dark for 3 minutes, and then read the plate. It is recommended to complete the plate reading within 10-30 minutes to obtain the best results.
4. Inter-plate internal reference: If inter-plate data comparison is required, it is recommended to set up two wells of positive controls without added compounds in all reaction plates as inter-plate internal references. The readings of each plate should first be corrected (normalization) using the inter-plate internal reference readings, and the corrected data should then be subjected to downstream processing and analysis.
5. Reaction plate: The level of fluorescence reading may cause obvious interference to the readings of adjacent wells, mainly due to the spillover of fluorescent signals. If necessary, opaque-bottom white plates or black plates can be used for experiments or verification.
6. Reagent mixing: It is not recommended to mix reagents from different batches, reagents that have been activated but not used up and stored, and reagents with significantly different storage conditions.
7. Aliquot storage: Dispense and store reaction reagents in accordance with the instruction manual to ensure the stability of the reagents.
