Immunoperoxidase labeling experimental steps - Database & Sql Blog Articles

Brand AVX TPSE226M035R0125 Low impedance tantalum capacitor - AVX 22

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

Probe current voltage pin, 420*4450 head diameter 5.0, over current and voltage pin

Huaqiang Square crystal original spot stock

1. Start by placing a wet tissue at the bottom of the slide box to create a humid environment. Take the slides with tissue sections from the frozen slicer and place them in the slide box, ensuring there are 6 slides on each side or in the wet box. Make sure the slides do not come into contact with one another.

2. While the slide is still moist, add PBS buffer to the section without spilling it off the slide.

3. Incubate the slides for 30 minutes at room temperature in PBS containing 0.25% hydrogen peroxide. Afterward, wash the slides three times with PBS to remove any residual reagents.

4. Dilute the first antibody at 4°C using a microcentrifuge at 13,500 g for 2 minutes. Ensure that 40 to 50 μl of the diluted antibody covers the tissue section properly.

5. Using a Pasteur pipette connected to a pump, gently blot the PBS from one end of the slide. Then, add the antibody solution from the opposite end, cover the slide with the humidification box, and incubate for 1 hour at room temperature.

6. Perform three washes with PBS, each lasting 5 minutes. During each wash, add fresh PBS from one end of the slide and aspirate the old buffer from the other end to ensure thorough rinsing.

7. Apply a specific bridged secondary antibody to the slide and incubate for 1 hour at room temperature. Afterward, wash the slides three times with PBS to remove any unbound antibodies.

8. Add the PAP complex to the slides and let it stand for 1 hour at room temperature. Follow this with three additional PBS washes to remove excess reagents.

9. Develop the DAB substrate solution for 2 to 5 minutes at room temperature. The exact development time may vary depending on your experimental experience and desired staining intensity.

10. Finally, place the slides in the slide box and store them in a dark area—do not freeze them. This helps preserve the integrity of the stained tissue for future analysis or documentation.