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0Firefly luciferase and Renilla luciferase detection systems are mostly used in traditional reporter gene detection systems, but the short half-life of the luminescent signal and poor stability, etc., affect the experimental results and use and cause inconvenience. In view of the shortcomings of traditional flash type luciferase, Sheng Bio has developed a new generation of glow type luciferase detection system with more convenient operation, longer half-life and better stability through painstaking research, suitable for samples such as mammalian cells, protoplasts, etc.
Fig 1: Schematic diagram of the detection principle of luciferase reporter gene detection
Comparison of flash type and glow type
1. Operation process
Fig 2: The flash type operation process is more complicated
Fig 3: Glow type operation process, complete detection in one step
2. Comparison of performance characteristics
|
Type |
Advantage |
Disadvantage |
|
flash type |
High sensitivity and strong signal (about 10 times the glow) |
Weak stability, half-life end Requires automated sampler for bulk screening |
|
glow type |
Good stability and long half-life Suitable for high-throughput screening, does not rely on automatic samplers, just pipette operation |
Low sensitivity, weak signal (about 1/10 of flash type) |
3. Application direction
4. Data Display
Fig 5. Fluorescence intensity and stability of Yeasen detection system are comparable to P* brand
Fig 6. The Yeasen detection system interacts with luciferase, and the luminescence quenching effect of luciferin is comparable to that of the P* brand.
Fig 7. Yeasen detection system emits light normally in common medium, and the difference in luminescence effect is smaller than that of P* brand.
FAQs
Q1：Optimum reaction temperature for dual luciferase reporter gene?
A：at room temperature (20-22°C). Reaction components such as cell lysate, substrate working solution, etc. need to be adjusted to room temperature, because the reaction rates of the two luciferases are affected by temperature. In order to ensure the consistency of the experiment, it is recommended to adjust the reaction room temperature to room temperature.
Q2：How to choose a dual luciferase reporter gene vector?
A：1) Firefly luciferase is recommended to choose pGL-3 or pGL-4 or a self-constructed vector. .
2) Renilla luciferase is recommended to select pRL-TK or pGL-4 vector or construct the corresponding vector by yourself. It is recommended to use medium-strength promoters (such as TK) rather than strong promoters (such as SV40, CMV), so as not to affect the expression of the main reporter gene, or to conduct experiments to find a suitable vector.
Q3：What is the appropriate ratio of firefly luciferase readings to Renilla luciferase readings?
A：It is recommended that the readings of control firefly and Renilla luciferase are close, or that the reading of Renilla luciferase is slightly higher than that of firefly luciferase. During the actual experiment, due to the inconsistent quality of the plasmids extracted from firefly luciferase and Renilla luciferase, and the different regulatory capabilities of the target regulatory elements to be detected, we recommend pre-experiment optimization of firefly luciferase. Co-transfection amount of plasmid and Renilla luciferase plasmid.
Ordering Information
|
Product Name |
Cat# |
Size |
|
Firefly Glo Luciferase Reporter Gene Assay Kit (Inquire) |
11404ES60/80 |
100/1000 T |
|
Dual Glo Luciferase Reporter Gene Assay Kit (Inquire) |
11405ES60/80 |
100/1000 T |
Related Products
|
Product Name |
Cat# |
Size |
|
Luciferase Reporter Gene Assay Kit (Inquire) |
11401ES76/80 |
500/1000 T |
|
Dual Luciferase Reporter Gene Assay Kit (Inquire) |
11402ES60/80 |
100/1000 T |
|
40902ES01/02 |
100/500 mg |
|
|
40901ES01/02 |
100/500 mg |
|
|
40802ES03 |
1.0 mL |
