Hello! I try to stain some synaptic proteins (synaptophysin, synapsin, SNAP-25) by immunofluorescence in synapses on mSOD1 mouse diaphragms, but I have faced 2 problems - first, sometimes the background fluorescence is high too much. Second, sometimes specific fluorescence of the proteins absents absolutely.
So I am wondering does anybody have a good, well-working protocol? Could you please share it with me?