When using the Alexa Fluor 647 Tyramide SuperBoost Kit, goat anti-mouse IgG (Cat. No. B40916), do I have to remove the 100 µL of tyramide working solution before adding 100 µL of reaction stop reagent (see step 3.2 and 3.3 in User Guide)?
It is not necessary to remove the 100 µL of tyramide working solution (step 3.2 in User Guide) before adding the 100 µL of reaction stop reagent (step 3.3 in User Guide; see link here: https://assets.thermofisher.com/TFS-Assets%2FLSG%2Fmanuals%2FMAN0015834_Tyramide_SuperBoost_Kits_UG.pdf).
You can just add it directly to the cell or tissue sample.
With a SuperBoost tyramide kit, I got excessive and non-specific labeling. What can I do to limit background and acquire a more localized labeling?
To limit background, we recommend performing a pre-blocking step with 3% H2O2 for 60 mins to inactivate endogenous peroxidases. To limit the localization of labeling, we recommend optimizing the final concentration of the primary and secondary antibodies and the dye-tyramide. You may also limit the incubation time of the dye-tyramide on the sample.