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What are luciferase assays used for?

What are luciferase assays used for?

A luciferase assay is used to determine if a protein can activate or repress the expression of a target gene.

How is luciferase activity measured in cells?

Cell debris is removed by microcentrifugation and luciferase activity is measured using a luminometer. Some luminometers directly inject the reagents into the cell lysate. Such automation allows the signal to be measured at a precise time following injection, which can increase the consistency of the results.

How do you lyse cells for luciferase assay?

Harvesting and Lysis of Cells

  1. iv. Transfer nonadherent cells to a 15-mL tube.
  2. v. Add 100 μL of Reporter Lysis Buffer to the 15-mL tube per each 106 nonadherent cells.
  3. vi. Resuspend the cells using a micropipettor to facilitate the disruption of cell clumps.
  4. vii. Transfer the lysed cells to a microcentrifuge tube.

What wavelength is luciferase assay?

approximately 550 – 570 nm
The luciferase protein from the firefly beetle (Photinus pyralis) is a monomeric enzyme that catalyzes the oxidation of luciferin while emitting light at approximately 550 – 570 nm.

What is a promoter assay?

Promoter analysis can be done by analyzing the expression levels of RNA. The assumption is that genes that have similar expression levels have similar transcriptional regulation control and common binding sites. This knowledge is used we have to find promoter regions on genome sequences.

What is dual luciferase reporter assay?

The Dual-Luciferase® Reporter (DLR™) Assay System provides an efficient means of performing two reporter assays. In the DLR™ Assay, the activities of firefly (Photinus pyralis) and Renilla (Renilla reniformis or sea pansy) luciferases are measured sequentially from a single sample.

What does luciferase mean?

Definition of luciferase : an enzyme that catalyzes the oxidation of luciferin.

What type of protein is luciferase?

Dinoflagellate luciferase is a multi-domain eukaryote protein, consisting of an N-terminal domain, and three catalytic domains, each of which preceded by a helical bundle domain.

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