Unfixed frozen samples were cut into 30-μm–thick sections and placed on glass slides. Dihydroethidium (10 μmol/l) was applied to each section, and then the section was incubated in a light-pro- tected humidified chamber at 37°C for 30 min. A fluores- cence image of DHE was ...
IHC sections were set on poly-l-lysine-coated glass slides, deparaffinized, and rehydrated in graded concentrations of ethanol, followed by heat-induced epitope retrieval. Endogenous peroxidase was blocked using methanol containing 3 % H2O2 for 15–30 min. Slides were blocked with BSA (Sigma) ...
T cells were obtained by gently grinding the thymus between the roughened glass of 2 microscope slides, and the cells were washed free with buffer. 20 μg of cellular protein (equivalent to 25,000 neutrophils) was analyzed by immunoblotting for Ym1 and neutrophil elastase (25) and for Ym1...