Immunohistochemistry for SULT1E1
Tumor and its corresponding surrounding tissues were embedded in
paraffin, serially sectioned at 5μM by an automated cryostat slicing
machine (Leica Bio systems). Sections were deparaffinized by baking at
60 c followed by xylene treatment, downgraded alcohol and water. Slides
were washed with PBST containing 1% casein for 10 minutes, tissue
sections were incubated with 5% casein for 30 minutes for preventing
non-specific binding followed by overnight incubation in primary
antibody SULT1E1 and HIF1αin 1% casein PBST, washed with 1% PBST and
incubated in 1% casein PBST containing secondary antibody for 1 hour,
washed with 1% PBST followed by water and stained with chromogenic
substrate DAB for 3 minutes and then washed with water. Slides were
fixed with mounting medium and observed under a microscope (Nikon,
Eclipse LV100, magnification 20X) to study the SULT1E1 expression and
localization.