Double immunofluorescence labeling when one fluorophore is amplified by TSA
I am trying to use immunofluorescence to label two proteins when one of the two fluorophores is amplified using a TSA kit (Invitrogen). Unfortunately, the second fluorophore (not amplified) is very dim compared to when it is imaged alone. I have applied the second fluorophore both after the TSA procedure and before and it doesn't seem to matter. Also, I block for at least an hour between protocols. Any suggestions?