I am doing NF-kB staining, but can not stain well inside the cell
(especially inside nuclei).  We use frozen section and now started to
try Saponin and proteinase K before the staining.  Some people in other
lab suggeted me to do overnight incubation for normal goat serum
blocking, 1st antibody and secondary antibody(anti-rabbit, fitC
conjugated).  I have never done overnight for blocking or secondary.
Doesn't it do harm for fluorescence?

Thank you in advance.


--
Masayuki Miyagishima, MD
University of Pittsburgh,
Department of Surgery