I am setting up an assay to detect LC3 expression in macrophages undergoing autophagy. Papers that I have read to date all seem to transiently transfect the cells with a GFP-LC3 construct prior to autophagy induction and then analyse by microscopy.
Has anyone out there used a fluorescent antibody to LC3 rather than doing a transfection? I thought that perhaps I could induce autophagy then add a fluorescent tagged LC3 antibody and then look by fluorescent microscopy (or confocal).
Thanks in advance