Dual Antigen Retrieval Method for Paraffin-Embedded Tissue Sections (Trypsin + Citric Acid)

2. Required reagents: Citric acid antigen retrieval solution (Maixin product number: MVS-0100/0101 or MVS-0066), trypsin antigen retrieval solution (Maixin product number: DIG-3008), distilled water, PBS buffer.

3. Operating steps:

(1) Deparaffinize and hydrate paraffin sections, rinse with tap water (it is recommended to remove tap water and draw a circle 3mm away from the tissue with an immunohistochemistry pen (Maixin product number: PEN-0002)), rinse with PBS 3×3 minutes;

(2) Remove PBS from the sections, add an appropriate amount of prepared working trypsin solution onto the sections, and place the sections in an incubation box for 15 minutes. Rinse with PBS 2×3 minutes. Rinse with tap water and soak in water for later use;

(3) Take an appropriate amount of diluted working citric acid antigen retrieval solution (800-1500mL) into the pressure cooker, place the pressure cooker on the induction cooker, and heat at high power until boiling;

(4) Adjust the power to medium (800-1000W), place the sections on a high-temperature resistant staining rack, put them into the boiling retrieval solution, cover the pot lid, fasten the pressure valve, and continue heating;

(5) When the pressure valve starts to release steam, time for 1.5-2 minutes, then remove the pressure cooker from the induction cooker. Allow it to cool naturally at room temperature for 10 minutes. Tap water can be sprayed on the outer wall of the pressure cooker to accelerate cooling of the retrieval solution inside. After cooling to room temperature, remove the sections, rinse thoroughly with tap water, rinse with PBS 2×3 minutes, and then proceed according to the operating steps of the relevant Maixin kit.

4. Precautions: For details, refer to the precautions for the antigen heat retrieval method and antigen enzyme digestion retrieval method for paraffin tissue sections.