20
SHIGEYOSHI OBA AND TAKEHIKO KOTI
Deparaffinization and rehydrate
.!.
PBS 5mins
.!.
Proteinase K 15mins at 37°C
PBS 5minsX3
.!.
Preincubate with TdT buffer RT 30mins
TdT reaction 37°C 2hrs
Tris HCl rinse 5minsX3
0.3% H20 2 RT 15mins
.!.
Rinse BrijlPBS 5minsX3
.!.
Normal goat IgG RT Ihr
.!.
HRP-conjugated goat anti-biotin RT 2hrs
.!.
Rinse BrijlPBS lOminsX4
.!.
DAB-H202 with CoCh and NiSOiNH4hS04 RT 5mins
.!.
Rinse distilled water> 5mins
Dehydration and mounting
Fig. 1. Paraffin block with biotin-16-dUTP