Immunoperoxidase labeling experimental steps - Database & Sql Blog Articles

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Electronic scale crystal oscillator 3.2*2.5mm 3225 16M (16.000MHZ) 12PF 10PPM 20PPM 30PPM

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1. Place the wet tissue on the bottom of the slide box to make the humidification box. Remove the slides containing the slices from the frozen slicer and place them in the slide box (6 pieces on each side), or in the wet box. (The slides should not touch each other).

2. When the slide is wet and not dry, add PBS to the slice (do not spill the slide).

3. Incubate for 30 min at room temperature in PBS containing 0.25% hydrogen peroxide and wash 3 times with PBS.

4. Dilute the first pellet at 4 °C with a microcentrifuge at 13 500 g for 2 min (40 to 50 μl of antibody per slide should cover the section).

5. Using a Pasteur pipette connected to the pump, blot the PBS on the slide at one end of the section and add the antibody from the other end, cover with a humidification box, and incubate for 1 h at room temperature.

6. Add PBS buffer 3 times (5 min/time), add new PBS buffer from one end of the section, and aspirate the old buffer from the other end.

7. Add a specific bridged secondary antibody to the slide and incubate for 1 h at room temperature. Wash 3 times with PBS.

8. Add PAP complex and let it stand for 1 h at room temperature. Wash 3 times with PBS.

9. The DAB substrate solution is developed for 2 to 5 minutes at room temperature. (The length of color development depends on experimental experience).

10. Place the sepals and fix them in a dark place in the slide box (not frozen).