Luciferase Assay Kit

Assay Kit for easy and efficient measurement of luciferase activity using cultured cells

• Utilized the properties of fire-fly luciferase which selectively reacts only with a specific substrate (luciferin)
• A brief experimental procedure to measure luminescence using transfected cell lysates
• The most representative experimental method among the reporter assays identifying various functions (gene regulation, promoter activity measurement, new drug screening, etc.) according to experimental conditions
• High sensitivity and low background value

Fire-fly luciferase is one of the reporter genes most commonly used for gene regulation or functional studies or for drug screening. The activity of luciferase can be measured easily and efficiently in cultured cells. Especially, fire-fly luciferase is easy to use because it selectively reacts only with specific substrate (luciferin). Luciferase Assay Kit will measure luciferin gene expression by the amount of luciferase expressed in each transfected cell by adding luciferin to each cell. Luciferin produced by transfected cell is oxidized under the condition of ATP and Mg2+ then emits light. At this time, luminescence is read by using luminometer. The product also has the advantage of being able to meet various experimental purposes. Even this makes it possible to get results in a short time using lysates of transfected cells. In this experiment, a vector containing a sequence in which β -Galactosidase is expressed is mainly used as an internal control to eliminate transfection efficiency depending on each cell during transfection. By using this vector, transfection rates with different values can be made equal and then gene expression values can be compared only. So it is a little easier to interpret.

  Applications

  Notice

The results of this Luciferase Assay Kit may differ depending on the storage method and storage temperature of each reagent used in the experiment. Therefore, please pay attention to the following points in the experiment. Luciferin solution should be kept free from exposure to light, avoided repeated freezing-thawing, and that activity may be reduced upon prolonged exposure to light. Also, basically it should be kept at -60°C or lower. Storage at -20°C or higher may reduce the activity and storage period of the reagent.

  Kit Contents

  Technical Data

Experimental results of Luciferase Assay Kit

Fig. 1. Data shows efficiency of Luciferase Assay Kit    
– Empty pCMV vector : over-expression vector backbone without cloning
– NFkB pCMV vector : over-expression vector overexpressing NFkB interacting with NFkB pGL3 vector
– NFkB pGL3 vector : reporter vector that interacts with NFkB and then produces luciferase

In this experiment, the pSV vector (including β-Galactosidase gene) was used to match the transfection value, and β-Galactosidase Assay was used to correct the results that may vary depending on the difference in transfection value. We used luciferase assay kit to check luminescence.

  Schematic Diagram

Fig. 1. Data shows efficiency of Luciferase Assay Kit    
– Empty pCMV vector : over-expression vector backbone without cloning
– NFkB pCMV vector : over-expression vector overexpressing NFkB interacting with NFkB pGL3 vector
– NFkB pGL3 vector : reporter vector that interacts with NFkB and then produces luciferase

In this experiment, the pSV vector (including β-Galactosidase gene) was used to match the transfection value, and β-Galactosidase Assay was used to correct the results that may vary depending on the difference in transfection value. We used luciferase assay kit to check luminescence.