I have very recently incorporated a HaloTag strategy for pulse-chase labeling in live cells into my project. I have checked a couple of protocols for live cell imaging using HaloTag ligands but the dilution (final concentration rarely mentioned), vehicle, duration of the pulse, washes, etc. vary widely. I guess they are optimized for different applications but I am quite confused and as a novice in the technology I don’t want to waste reagent (and cells… and time…). Does anybody have any guidelines and tricks that work?
I have HaloTag ligand conjugated to Janelia Fluor dyes as well as AlexaFluor660.
Thanks in Advance!