[Histonet] Problems with Gold-immunolabeling and silver enhancement in brain tissue
I'm trying to perform silver enhancement of my gold
labeled secondary antibody (1.4nm gold) in an
immunocytochemistry (ICC) protocol in rodent brain
tissue. I know that the primary antibody and initial
steps are working because when I use an
avidin-biotinylated peroxidase kit to visualize a
biotinylated secondary antibody (as opposed to using
the gold conjugated secondary antibody)I get great
labeling of my antigen. I've done a dot blot with the
gold labeled antibody and I seem to get silver
enhancement of particles only in the dot blot with the
gold-labeled antibody (as opposed to unlabeled
antibodies)- so I'm pretty sure there are gold
particles attached to the IgG.
Are there any obvious differences that need to be
considered when working with a gold-conjugated
antibody as opposed to antibodies that are
biotinylated or conjugated to enzymes??
Any insights would be greatly appreciated..
Department of Physiology and Biophysics
University of Illinois
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