I'm trying to quantify the expression of a protein in spinal cord . I have obtained significant differences in western blot between two groups of animals, but quantification after fluorescent immunostaining (confocal microscopy) doesn't show so evident differences.
Antibody used is the same in WB and IF. Slices were processed in the same conditions with the 2 groups of animals on the same slide.
Maybe the antigen conformation could be involved, but staining is ok in WB and IF!?
If you have any opinion on this problem and maybe solutions...
Thank you very much for your help