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After permeabilization with PBS containing 0.1% Triton X-100 for 10 minutes sections were blocked for 1 hour with PBT (0.15% BSA, 0.1% Tween 20 in PBS).
After pretreated with 0.3% hydrogen peroxide in PBS for 15 minutes, sections were preincubated for 1 hour in blocking solution (0.1 M phosphate buffer, 0.25% Triton X-100, and 5% normal goat serum).
After leaving at room temperature for 20 minutes, sections were immersed in phosphate-buffered saline (PBS).
After cooling at room temperature for 20 minutes, sections were rinsed in deionized water.
After washing with PBS (3 × 5 minutes), sections were incubated with biotinylated secondary antibodies for 30 minutes.
After incubation of each antibody for 30 minutes, sections were rinsed and incubated with the appropriate secondary antibodies (DAKO).
After deparaffinization in xylene for 30 minutes, sections were rehydrated through a gradient with decreasing proportions of ethanol.
After washing with PBS, 3%H2O2O2 treated for 10 minutes, sections were blocked with 10%% serum for 20 minutes at room temperature.
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CEO of Professional Science Editing for Scientists @ prosciediting.com