Hello all.

Does anyone know if the biotin on a biotinylated primary antibody interpheres with a secondary antibody? I am not getting any staining after indirectly labeling a biotynilated antibody and wonder if that is a possibility.

Thank you.

Why you didn't use the streptavdin conjugated secondary antibody if you use the biotinylated primary antibody.

I had the secondary antibody with the fluorochrome I wanted so I decided to try it. I did try streptavidin but the staining was too "dirty" to see anything. Also, I haven't used this method without an amplification kit, like ABC, so I am not sure the signal will be good enough.