Contents

Storage 

Store at -80 ℃. Component A, B and D is stable for at least 3 months at -20 ℃ and 6 months at -80 ℃. Component C and E is stable for at least 6 months at -20 ℃ in dark. Firefly luciferase working solution (B + C) is stable for one month at -20 ℃ and 3 months at -80 ℃, and do not freeze and thaw repeatedly, it is recommended to pack in small batches. Renilla Luciferase Assay solution(D + E) should be freshly prepared and used on the same day.

Description

Firefly & Renilla Dual Luciferase Assay Kit provides an effective means for double reporter gene detection. In DLR detection, the activity of Firefly luciferase and Renilla luciferase can be detected in a single sample in turn. First, use luciferin as substrate to detect Firefly luciferase, and then use coelenterazine as substrate to detect Renilla luciferase, and when luciferase substrate is added subsequently, substances that inhibit luciferase catalytic luciferin luminescence are added at the same time, so that the subsequent detection only detects luciferase activity of Renilla luciferase and realizes the detection of double luciferase reporter gene. Luciferase and its substrate, a bioluminescent system, can detect gene expression very sensitively and efficiently. Usually, the transcriptional regulatory element or 5 'promoter region of the gene of interest is cloned upstream of luciferase, or the 3' - UTR region is cloned downstream of luciferase, etc. to construct a reporter gene plasmid, and then transfect into the cells. After the cells are treated with appropriate drugs, the cells are split to determine the luciferase activity. Through the activity of luciferase, we can judge the transcriptional regulation of target gene by drug treatment. Renilla luciferase was used as an internal parameter of transfection efficiency to eliminate the difference of cell number and transfection efficiency.

Firefly luciferase, a monomeric 62,000 Dalton protein, catalyzes ATP-dependent D-luciferin oxidation to oxyluciferin to produce light. Renilla luciferase has been used as a reporter gene for studying gene regulation and function in vitro and in vivo. It commonly is used in multiplex transcriptional reporter assays or as a normalizing transfection control for firefly luciferase assays. Renilla luciferase, a monomeric 36,000 Dalton protein, catalyzes coelenterazine oxidation by oxygen to produce light. This kit has the characteristics of rapid detection, high sensitivity, wide detection range and no interference of endogenous activity.

Notes

1. In order to obtain the best measurement effect, the time from the mixture of sample and reagent to the time before determination should be controlled alike as much as possible when using the single tube chemiluminescent apparatus; when using the multi-functional fluorescent enzyme reader with chemiluminescent function, it is advisable to add all the samples first, and then add the fluorescent luciferase detection reagent uniformly. 

2. Because the temperature has an effect on the enzyme reaction, the sample and reagent should be determined after reaching the room temperature. 

3. For your safety and health, please wear lab coats and disposable gloves.