Dual luciferase reporter assay system instructions for use of products e1910 and e1960. For an overview and information on the development of the dualglo luciferase assay system, see sections 6. Luciferase assay expected range of raw reading values. Development and characterization of a novel luciferase. For manual luminometers, the substrate solution is mixed by hand with the. Cells were harvested 48 h post transfection and subjected to the luciferase assay following muthumanis protocol with the protein concentration of the lysate as a reference 28. Luciferase reporter assay in drosophila and mammalian tissue. Unless otherwise expressly authorized in writing, products are supplied for research use. Luciferase assay expected range of raw reading values hi there, i am performing luciferase assays with chromaglo luciferase assay system from promega using the mutlilabel counter victor3. Nanoglo luciferase assay system technical manual, tm369.
The protocol below is for manual assay using a singletube luminometer. Firefly luciferase vectors for use with the oneglo ex luciferase assay system. The dualluciferase reporter dlr assay system provides an efficient. Prepare a mix of 20ul of dualglo luciferase reagent and 20ul serumfree media for each well. Renilla luciferase has been used as a reporter gene for studying gene regulation and function in vitro and in vivo1, 2. Dual luciferase and dual luciferase assay protocols dual luciferase reporterassay and dual luciferase reporter assay systems instructions for use of products e1910, e1960 and e1980. Dualluciferaser reporter assay system technical manual tm040. Luciferase reporter assays allow you to investigate promoter activity by measuring light output from luciferase enzyme that is expressed under the control of your promoter of interest. Luciferase genes vary in stability, light output and spectral characteristics, and reporter kits are available with different luciferases for both single and dualspectral analyses. Product components and storage conditions product size cat.
Usa toll free in usa 8003569526 6082744330 fax 608277 2516. Dualluciferase reporter assay system promega corporation. Use a standard protocol to transfect mammalian cells with a firefly luciferase plasmid. Firefly luciferase is widely used as a reporter for studying gene regulation and function, and for pharmaceutical screening1, 2.
Thaw the nanoglo luciferase assay buffer if stored at 20c but do not exceed 25c. The linear range of the firefly luciferase assay extends over 8 orders of magnitude of. Development of a cytotoxicity assay based on transient expression of different luciferases in cytosol. Measuring firefly and renilla luciferase activities in. The luciferase reporter assay is commonly used as a tool to study gene expression at the transcriptional level.
Dualluciferase reporter assay system harvard university. Luciferase reporter gene assays are one of the most common. To see articles that cite the use of the dualglo luciferase assay system, visit. It has broad applications across various fields of cell and molecular biology wherever you want to measure or track expression of a cloned gene. Firefly luciferase is widely used as a reporter for studying gene. Luciferase reporter assay kit university of michigan. Luciferase reporter assay kit ii renilla promocell. It is widely used because it is convenient, relatively inexpensive, and gives quantitative measurements instantaneously. The dualglo luciferase assay system is designed to work in growth media commonly used for mammalian cells with or without added serum. In this protocol, cells transfected with a luciferase reporter plasmid are lysed using a. Introduction firefly beetle photinus pyralis luciferase is one of the most popular reporter. Toll free in usa 8003569526 phone 6082744330 fax 6082772516.
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