Since the discovery of green fluorescent protein, scientists have been finding new ways to use fluorescent proteins (FPs) as visual reporters for their experiments.They have been used to monitor protein localization, protein interactions, and more. And, the number of FPs available keeps increasing as scientists identify and modify fluorescent proteins. In this section of the blog we’ll cover some of the basics of working with FPs, and the many tools for imaging, biosensors, FRET, and more. We’ll also cover luminescence and non-protein fluorophores.
From the basics of fluorescent proteins (FP) to how to choose one, this section is a great place to get started with using fluorescent proteins in your experiments.
Fluorescent biosensors can be used to monitor a process or detect a molecule. Biosensor activity can be detected using microscopy or by measuring a change in fluorescence intensity or wavelength. Find some examples of fluorescent biosensors with plasmids available at Addgene in this section.
In this section of the blog, we describe different fluorescent protein and microscopy techniques you can use for imaging. There are many ways to use fluorescent proteins for imaging such as in vivo imaging or multi-color imaging, and more.
FRET, which stands for Förster Resonance Energy Transfer, measures the transfer of fluorescence of energy from an excited donor fluorophore to an acceptor fluorophore. This method helps scientists monitor protein interactions and more!
There are many ways to fuse a fluorescent protein to your protein of interest. From deciding where to fuse the fluorescent protein to what fluorescent protein you will use to using scaffolds, there are many options!
Because of their visual readouts, fluorescent proteins are great tools to identify where your protein of interest is located in the cell. Knowing the location of your protein can help scientists better understand its function. Localization studies can be done with more than one fluorescent protein so that more than one protein of interest can be tracked at the same time.
While luminescence is not a fluorescence readout, it can still be used for similar studies. Luminescence is a characteristic of the luciferase enzyme which emits photons during a chemical reaction. For example, the firefly luciferase converts luciferin to oxyluciferin and releases light during that process.
Proteins aren’t the only things that can be used for fluorescence. In addition to fluorescent proteins, there are chemical compounds that can emit light when excited. These are often used for staining.
Planning to use specific fluorescent protein plasmids for your experiment? This section features fluorescent protein plasmid tools developed by Addgene depositors not covered in the above sections.