Luciferase how does it work

Gong, D. Journal of Biological Chemistry, 8 , — Hampf, M. A protocol for combined Photinus and Renilla luciferase quantification compatible with protein assays. Analytical Biochemistry, 1 , 94— He, M. Matta, H. Development and characterization of a novel luciferase based cytotoxicity assay. Scientific Reports, 8.

Min, J. Journal of Clinical Medicine, 9 8 , Nakajima, Y. Multicolor luciferase assay system: one-step monitoring of multiple gene expressions with a single substrate. BioTechniques, 38 6 , — Smale, S. Luciferase assay. Cold Spring Harbor Protocols, 5 , pdb. What is a Luciferase Reporter Assay? In this protocol, cells transfected with a luciferase reporter plasmid are lysed using a detergent-containing buffer. Cell debris is removed by microcentrifugation and luciferase activity is measured using a luminometer.

Some luminometers directly inject the reagents into the cell lysate. Roundtable Discussion. Transitioning high-content assays to 3D: Scientific opportunities and imaging challenges.

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I think it measures transcription actually because what you do is that you insert the luciferase gene wich id the enzyme that will aid the conversion of luciferin wich is a protein. I guess the Luciferase translation is necessary for the measurement. However, it does not mean that your gene of interest has been translated into protein just that its promoter is less or more active. Translation in addition to transcription is certainly needed to obtain the luciferase catalyse the reaction.

However, the variable of interest would likely be transcription, based on the assumption that increased transcripts results in increased protein rather than a specific increase in production of the translation machinery given the same number of transcripts to increase protein. We basically compare the effect of the reporter gene with and without the miRNA binding sites.

So both the constructs have the same promoter and the difference in the enzyme activity would have to be at a translational level. So the variable of interest can be both at a transcriptional level as well as the translational … in this case.

Facebook Twitter LinkedIn More. Written by Farida Khan. Tammy Ferguson on October 19, at pm.