Signal Stability Detection: THP1-Dual cells at a concentration of 5×10⁴/ml were seeded into a 12-well cell culture plate. After stimulation with 2'3'-cGAMP to a final concentration of 3mg/ml for 24 hours, the supernatant was collected for enzyme activity analysis. Detection was carried out according to the kit instructions, with 50ml of detection reagent added to 20ml of supernatant. After mixing, fluorescence readings were taken at different time points. (The diamond line represents the detection results of the UA-Luc 4 Luci/Gaussia kit, while the black dot line represents the detection results of other brand products. The samples and their quantities are the same.)
UA-Luc 4 Lucia/Gaussia Luminescence Detection Kit
UA-Luc 4 Lucia/Gaussia Luminescence Detection Kit
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$209.00 SGD
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Product Details
Product Details
Product Specification
| Synonyms | 4 Luci/Gaussia荧光素酶检测试剂盒 |
| Stability & Storage | The 4 Luci/Gaussia Luciferase Assay Kit can be stored at -20°C or below for stable performance for up to one year. The prepared 1x reaction solution can be aliquoted and stored at -20°C in the dark for one month or at 4°C for 48 hours. Avoid repeated freeze-thaw cycles. |
Background
The Luciferase-based reporter gene system is widely used in the study of intracellular signaling pathways, transcription factor regulation, receptor function, and high-throughput drug screening. The expression of intracellular luciferase can be quantitatively detected. The working principle is that luciferase catalyzes the substrate, causing a transformation that produces spontaneous cold light. The intensity of the light is directly proportional to the amount of luciferase.
The UA-Luc 4 Luci/Gaussia Luciferase Detection Kit is a homogeneous ready-for-use reagent that completes the detection through the simple steps of “add-mix-detect.” This kit is used to detect the luciferase activity in the culture medium of cells transfected with secretory Luci/Gaussia luciferase reporter genes. There is no need to lyse the cells, which can be used for other analyses. Compared with similar products from foreign brands, the detection sensitivity is about one order of magnitude higher, making it particularly suitable for the analysis of cells with low enzyme activity and the screening of inhibitory compounds.
Components
Size |
Buffer |
Substrate |
Tests in 96-well plates |
Tests in 384-well plates |
500 tests |
25 ml |
250 ml |
500 |
2,500 |
5*500 tests |
5x 25 ml |
5 x 250 ml |
2,500 |
12,500 |
Protocol
1. According to the experimental design, cells expressing secretory Luci/Gaussia luciferase reporter genes are seeded into 96-well or 384-well reaction plates.
2. Cells are treated with compounds and stimulated with the reporter gene as required by the experiment.
3. At an appropriate time after reporter gene stimulation, such as 24–48 hours, collect the cell supernatant for luciferase activity analysis.
4. Before the experiment, equilibrate the UA-Luc 4 Luci/Gaussia Luciferase Detection Reagent to room temperature and gently mix.
5. Prepare the corresponding volume of 1x reaction solution according to the required amount. The substrate in the kit is 100x and should be diluted to 1x reaction solution with buffer before use.
6. Add 20 µl of the detection sample or control sample to each well of the 96-well white opaque plate, followed by the addition of 50 µl of detection reagent to each well.
7. Vibrate on a plate shaker at low speed for 10 seconds.
8. Immediately read and record the fluorescence signal on a fluorescence plate reader.
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Bioactivity
