I would like to photoactivate, via laser, a protein with GFP tagged to it (inside a solution) and then visualize the activated GFP's fluorescence on a non-denaturing polyacrylamide gel. An alternative would be to somehow probe only photoactivated protein by either using an antibody that is specific to the photoactivated form of GFP, which does not seem to exist, or through another method that I am not aware of. Can this be done? Any suggestions and tips would be appreciated. Thanks!
It is impossible to get a antibody that is specific to photoactivated form of GFP becuase GFP has the same conformation to photoactivated GFP. I have not visualized the GFP protein on polyacrylamide gel. In my experience, detection of DNA or RNA on gel is very sensitive. Theoretically, it is highly possible to visualize GFP on gel since GFP is relatively stable in nature. The only concern is that the loading buffer contains SDS disrupting the normal structure of GFP. So my suggestion is performing buffer exchange to the gel before scanning. This is something similar to the protein refolding step in protein purification.
Hypothetically, your GFP should be excitable in a native PAGE much like you suggested.
I found a protocol that uses that exact approach to find GFP-tagged proteasome. Check it out and see if that helps!
https://link.springer.com/protocol/10.1007%2F978-1-61779-474-2_23
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