Luciferase Assays
Reporter gene technologies are excellent tools to study gene expression, regulation as well as complex cellular processes such as gene networks. Reporter systems based on luciferase represent a highly sensitive, simple and flexible method which is especially suited for high throughput screenings.
The Amaxa® Nucleofector® provides easy and reproducibly efficient transfection of almost any cell type in single reactions or up to 96-wells at a time. In combination with Promega's luciferase reporter technology it leads to rapid and high expression even in scientifically most relevant primary cells. For transfection optimization luciferase reporter technology is a well suited alternative to GFP, especially for high throughput applications.
Benefit from
- Luciferase expression within just a few hours
- Working with primary cells or cell lines
- Easy optimization of transfection conditions
- Quantitative analysis using standard plate readers
Nucleofection® enables luciferase assays in primary cells.
Human Mesenchymal Stem Cells (HMSC) and Human Umbilical Vein Endothelial Cells (HUVEC) were transfected by Nucleofection® with a firefly luciferase expression plasmid. Luciferase expression was analyzed 6, 24 and 48 hours post Nucleofection® by Dual-Glo Assay (Promega). The 24 h value was set to 1.
Multiplexing bioluminescence with cell viability assays for rapid transfection optimization.
Jurkat cells [ATCC TIB-152] were transfected by Nucleofection® using a firefly luciferase reporter vector. Reporter gene expression and toxicity were analyzed 24 hours post Nucleofection®. Each transfection was characterized by measuring cell viability using the CellTiter-Fluor Assay (Promega) and then multiplexing it with the luminescence reporter assay (One-Glo Luciferase Assay, Promega). Viability of nontransfected control cells was set to 100%. (Data kindly provided by Promega Corp.)
