Histonetters,

I am trying to detect a protein in brain tissue using immunocytochemistry.
>From northern and western blots we know that the RNA is made and is 
translated into protein, now we want a more precise localization - it should
be nuclear but we are not sure in which cell type(s) it is expressed.

I have used a biotinylated IgG followed by the ABC kit from Vector which is 
a peroxidase reaction using DAB. No protein was detected although grossly,
the tissue had more of a yellowish tint compared to negative controls. Also,
we have a positive control which uses the same tissue preparation, i.e.
fixation, cryosectioning, etc. One current theory is that the protein is      
present in many cell types but at a low level of expression.

What would you say is the most sensitive immuno-detection method available?
Would fluorescence be better?

Thanks in advance
judy


Judy Trogadis
Eye Research Institute and
University of Toronto
Toronto Hospital, Western Div.
399 Bathurst  St.
Toronto, Canada M5T 2S8

phone: 416-603-5088
Fax:   416-603-5126
email: judy@playfair.utoronto.ca