I would like to use them as controls. I am studying the dynamics of receptors on the cell surface using SMI TIRF microscopy. I am currently using CHO cells and I would like to use constructs as controls with tunable dimerization affinities and which work with a halo tag and in CHO cells. I was thinking of either two different proteins (or a WT and mutant) that have well characterized/different on/off rates, or some sort of constructs with tunable/inducible dimerization.
I am unable to observe changes in dynamics between my protein of interest (VEGFR2) and its mutant which should have less dimerization, or changes in dimerization with ligand stimulation. I am not sure because it is being expressed in CHO cells, or due to the Halo Tag label. Could anyone please recommend me a construct which has been previously used for similar purposes? preferably not constructs that are not all either monomers or dimerized, but are able to switch back and forth, and can be labelled with little problem or are already stably expressed with Halo tag in a cell line.
Any suggestions would be greatly appreciated! I have been struggling with my current proteins for a while now.
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