[Histonet] double/triple staining with streptavidin-Alexa Fluor

[Yves Heremans]

Dear HistoNetters,

Is there a way to perform double (or triple) staining using secondary, biotinylated antibodies with streptavidin-Alexa Fluor conjugates, for instance, anti-rabbit, biotinylated secondary antibody A, detected with streptavidin-AF488 in combination with anti-mouse, biotinylated secondary antibody B, detected with streptavidin-AF555, preventing cross-reaction ? Will sequential staining (i.e. first anti-rabbit and SA488, then anti-mouse and SA555) saturate all biotin binding sites from the first step ?

Yves