GFP Transfection Efficiency Assay
– using the NucleoCounter® NC-3000™
- Fixed protocol – no settings needed
- Fully automated – just load sample and press run
- High specificity – assess transection at single cell level
- Results provide image, scatter plot and GFP histogram
- Results are precise and reproducible
Run 8 samples and analyze in under 20 minutes
Figure 2. Just add dye to your sample, incubate for 15 minutes, add another dye, load 8 chamber slide, and press RUN.
The introduction and expression of foreign genes in eukaryotic cells is a technique widely used for investigating gene regulation and function. It is often required that the efficiency of introducing the new gene into the cells – transfection efficiency – is determined for either process optimization or for the accurate interpretation of down-stream results. A quick and easy method for determining transfection efficiency is to use the Green Fluorescent Protein (GFP) as a reporter. Using an appropriate promoter, GFP can be expressed in the cells by itself or attached to the protein of interest as a fusion protein. Using GFP as a reporter, the transfection efficiency can easily be determined as the percentage of cells expressing GFP in the entire cell population.
Visualize where cells fall on a scatter plot
The NucleoCounter® NC-3000™ is an image cytometer allowing users to visualize where cells fall on the scatter plot. This can be at the single cell level to set very tight thresholds between positively stained and negatively stained cells, as well as for groups of cells to investigate the staining pattern for specific cell populations.