I am struggling to get my LC3-I and LC3-II bands to separate on my western blot. I can see there is a difference in the protein levels but I'm not getting any distinction between the two.

I am currently collecting lysates with RIPA buffer and running them on a 15% SDS gel (I have also tried 12% and 18%) in Tris-Glycine buffer at 90V for up to 2h. I am then transferring to 0.2um PVDF membrane using a BioRad TransBlot Turbo.

I would like to be able to get a clear separation between these two bands so my analysis is a lot more reliable.

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