The V9 clone for vimentin from Boehringer-Mannheim has worked well for me
on pf-fixed sections.
deparaffinize with xylene, rehydrate thru alc series to water.
TBS rinse
block with 5% NGS in TBS for 30-60 min at room temp
apply V9 at 1:100 in 1% NGS overnite at 4 degrees
tbs rinses
secondary is goat antimouse IgG linked to fluorescine (fitc) at 1:200 in
tbs for 45 min at 37 degrees (Cappell)
rinse 3X with tbs for 10 min each
mount with antiquench mounting medium from Shandon
enjoy under blue light optics
alternatively, use V9 at 1:1000 and use standard immuno kit (ABC) like the
one from Vector
Good luck,
Lori DeGiorgio
Cornell med at Burke Med Res Inst
ldegiorgio at aol.com