Hi Kristóf!

I can't give you an exact response since your protein is very specific.

However, I can provide you some information based on my personal experience: once I used GFP antibody to perfom a Western Blot to see if HEK293 cells transfected with a DNA construct that contained an antibody fragment and a recoded GFP sequence expressed properly the construct (to check if the sequence was properly transcripted and translated into protein).

Results were good, because at least in the cell lisates we could see the band in the predicted molecular weight of the fusion protein.

I think the important thing here is to use proper denaturing conditions (SDS, lysis buffer and heat well the sample) to ensure that the protein unfolds properly and expose the epitopes to its surface.

What technique/s will you perform with the GFP antibody? Western, FACS...?

Also, what is the brand of the Antibody (mine I think it was Sigma Aldrich)

Hope it helps!

Best regards

Francesc

I don't know for your protein but if the primary sequence is concerved and your antibody recognize a linear epitope then it should be OK exept if the epitope is right at the junction of the permutation. so maybe a polyclonal antibody would be better. be carreful; the iGluSnFR protein has been mutageneized so some epitopes can be lost (some monoclonal antibodies against GFP do not recognize eGFP )

looking at Nature Methods volume 20, pages 925–934 (2023) figure 4 they did imaging of iGluSnFR with chicken anti GFP abcam ab13970....