HEK-293T and HeLa cells were seeded in 96-well plates and when 70−80 % confluent were transfected with either the NF-KB-Luciferase reporter plasmid (pNF-KB-Luc) or the IFN-stimulated response element-Luciferase reporter plasmid (pISRE-Luc) and a plasmid constitutively expressing Renilla Luciferase (pTK-RL). Following stimulation, cells were lysed in passive lysis buffer (Promega) and Firefly luciferase and Renilla luciferase were measured with Firefly luciferase substrate and Renilla luciferase substrate (Nanolight Technology) using a FLUOstar luminometer (BMG). The Firefly Luciferase activity was normalised to the Renilla Luciferase activity first, and then data were normalised to non-stimulated EV group. At least three independent (technical replicates) measurements were taken per condition per experiment.
Renilla luciferase substrate
Manufactured by Nanolight Technology
Renilla luciferase substrate is a reagent used to measure the activity of the Renilla luciferase enzyme. Renilla luciferase is a bioluminescent reporter that catalyzes the oxidation of coelenterazine, resulting in the emission of light. The substrate provides the necessary coelenterazine for this reaction, allowing for the quantification of Renilla luciferase activity.
Automatically generated - may contain errors
Lab products found in correlation
2 protocols using renilla luciferase substrate
1
Dual-Luciferase Reporter Assay for NF-kB and ISRE
2
Dual-Luciferase Reporter Assay for NF-kB and ISRE
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
HEK-293T and HeLa cells were seeded in 96-well plates and when 70−80 % confluent were transfected with either the NF-KB-Luciferase reporter plasmid (pNF-KB-Luc) or the IFN-stimulated response element-Luciferase reporter plasmid (pISRE-Luc) and a plasmid constitutively expressing Renilla Luciferase (pTK-RL). Following stimulation, cells were lysed in passive lysis buffer (Promega) and Firefly luciferase and Renilla luciferase were measured with Firefly luciferase substrate and Renilla luciferase substrate (Nanolight Technology) using a FLUOstar luminometer (BMG). The Firefly Luciferase activity was normalised to the Renilla Luciferase activity first, and then data were normalised to non-stimulated EV group. At least three independent (technical replicates) measurements were taken per condition per experiment.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!