Signal Stability Test of One-luc Luciferase Assay Kit: HEK293 cells were seeded into 96-well cell culture plates; after 5 hours, luciferase expression plasmids were transfected into the cells; 48 hours post-transfection,
the cells were taken out and equilibrated at room temperature for 10 minutes, then One-luc Luciferase Assay Reagent (equilibrated to room temperature) was added, mixed well, and the fluorescence
values were continuously read at different time points. The comparative reagent is a similar product from a foreign brand. The values in the figure represent the average of two replicates.
UA-Glo® One-luc Luciferase Assay System
UA-Glo® One-luc Luciferase Assay System
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Product Details
Product Details
Product Specification
| Synonyms | One-luc Luciferase Detection Kit |
| Stability & Storage |
The One-luc Luciferase Assay Kit remains stable in performance for at least 1 year when stored at 20°C or below. For reagents that cannot be used up in one go after opening the bottle, it is recommended to aliquot them and store at -20°C. It is advised that the number of freeze-thaw cycles does not exceed 3, and the time of each placement at room temperature should not exceed 2 hours. For short-term continuous use, the reagent can be stored in a 4°C refrigerator for no more than 5 days. |
Background
The luciferase-based reporter gene system is widely used in studies on intracellular signaling pathways, transcription factor regulation, receptor functions, high-throughput drug screening, etc. The expression of intracellular luciferase can be quantitatively detected. Its working principle is that in the presence of the substrate luciferin, ATP, and molecular oxygen, luciferase can oxidize luciferin into oxyluciferin, thereby producing spontaneous cold light. When excess luciferin and ATP are present, the intensity of the luminescence is positively correlated with the amount of luciferase.
The UA-Glo homogeneous ready-to-use stable luciferase detection kit is used for the quantitative detection of luciferase content expressed in cells. This reagent features high signal-to-noise ratio, good repeatability, and good stability. The ready-to-use formula eliminates the experimental steps of lysis followed by detection, thereby further reducing errors caused by frequent sample addition. The half-life of the detection signal of the stable luciferase detection kit can generally reach 2 hours, making it particularly suitable for high-throughput sample detection.
The One-luc luciferase detection kit has better chemical stability. When the detection reagent is stored at 4°C for 5 days, the detection reaction signal only decreases by about 15%, which is convenient for customers who need to use the kit repeatedly within a short period of time. Its performance is similar to that of the foreign ONE-Glo product.
Components
ATP, luciferin, and buffer solution are mixed and filled into 10 ml or 100 ml brown bottles, with specifications as follows:
|
Specification |
Detectable number of wells in 96-well plates |
Detectable number of wells in 384-well plates |
|
10 ml |
100 wells |
500 wells |
|
100 ml |
1,000 wells |
5,000 wells |
|
10X100 ml |
10,000 wells |
50,000 wells |
Protocol
1. Before the experiment, equilibrate the One-luc luciferase assay reagent to room temperature and mix gently by shaking.
2. Equilibrate the cell plate to be tested (white transparent-bottom or black bottom plate) at room temperature for 10 minutes.
3. Add an equal volume of the assay reagent to each culture well as the cell culture medium.
4. Shake on a plate shaker for 1 minute, then incubate the sample plate at room temperature in the dark for 5-8 minutes.
5. Read and record the fluorescence signal on a luminescence microplate reader.
Picture
Picture
Bioactivity
HEK293 cells were seeded into 6-well cell culture plates and transfected with luciferase expression plasmids; 48 hours post-transfection, the cells were digested with enzymes, counted, and then aliquoted into 96-well assay plates with the number of cells per well as shown in the table. Subsequently, One-luc Assay Reagent (equilibrated to room temperature) was added for detection, with untransfected cells as controls. The fluorescence values were continuously read at different time points, and the reading ratios were calculated. The comparative
reagent is a similar product from a well-known foreign brand. The values in the table are the average of two replicates.
