We have recently published a new protocol in STAR protocols, a journal by Cell Press. This protocol provides an in-depth, step-by-step protocol describing how to perform single-molecule Fluorescence Recovery After Photobleaching (smFRAP). This technique was developed as a method to determine the relative concentrations of nuclear envelope transmembrane proteins (NETs) on the inner and outer nuclear envelope.
This technique is particularly powerful as it provides the ability to determine not only the concentrations of the NETs on the discrete envelopes, but also the diffusion coefficient of the discrete NETs. This enables researchers to determine how quickly a NET is diffusing on the nuclear envelope, both on the inner and outer leaflet. This technique may also be expanded to determine import and export rates of discreet NETs and is currently the only technique capable of doing so.
The article is titled Protocol for single-molecule fluorescencerecovery after photobleaching microscopy toanalyze the dynamics and spatial locations ofnuclear transmembrane proteins in live cells. It is published by STAR protocols.
