Formalin fixation of cells or tissues causes cross-linking between proteins and between proteins and nucleic acids, which masks antigenic epitopes and impairs the results of immunohistochemical staining. Selecting a suitable antigen retrieval solution and adopting a heat-induced retrieval method can break these cross-links and fully expose antigenic epitopes to achieve the purpose of antigen retrieval. Meanwhile, the deparaffinization reagent components contained in this product can separate paraffin from tissues in sections, thus completing the deparaffinization and hydration processes simultaneously during heat-induced antigen retrieval.
Paraffin Section Deparaffinization Antigen Retrieval Solution, also known as One-step Deparaffinization/Antigen Retrieval Solution, Deparaffinization Heat Retrieval Solution and Deparaffinization Retrieval Solution, is prepared from Tris, EDTA, deparaffinization reagents, PC300 and deionized water. It should be diluted 20-fold with deionized water and adjusted to pH 6.0 before use. This product breaks the limitations of traditional paraffin section processing on experiments and eliminates the need for fume hoods and traditional deparaffinization jars. It integrates the traditional procedures of 2-3 rounds of xylene deparaffinization, 3-5 rounds of gradient ethanol hydration and antigen retrieval into a single solution, effectively shortening the operation time and simplifying tedious steps. At the same time, it reduces variables in the operation and improves the stability of subsequent staining. Adopting a new environmentally friendly technology, this product not only minimizes harm to the human body but also causes no environmental pollution. This reagent is for research use only and not intended for clinical diagnosis or any other applications.
Materials to Be Prepared by the User
1. Graded ethanol, double-distilled water or deionized water, immunostaining wash buffer
2. Heating equipment (microwave oven, water bath, autoclave, etc.)
3. Formalin-fixed, paraffin-embedded (FFPE) tissue sections
Operating Procedures (For Reference Only)
1. Prepare Deparaffinization Antigen Retrieval Working Solution: Paraffin Section Deparaffinization Antigen Retrieval Solution (20×, Low pH) is a 20-fold concentrated solution with alkaline pH. Dilute it 19-fold with deionized water to prepare the working solution before the experiment. Example: Take 25 ml of Paraffin Section Deparaffinization Antigen Retrieval Solution (20×, Low pH) and add deionized water to make the final volume up to 500 ml, and adjust the pH to 6.0.
2. Deparaffinization, Hydration and Antigen Retrieval:
(1) Place the deparaffinization antigen retrieval working solution in a retrieval jar and heat it to boiling using heating equipment.
(2) Immerse the sections in the boiling working solution (metal slide racks are not allowed to ensure the pH of the retrieval solution) and ensure the tissue on the sections is completely submerged.
(3) Continue heating on low to medium heat for 15-30 minutes. Do not let the tissue dry out during this process. Remove the retrieval jar from the heat source and allow it to cool to room temperature naturally.
(4) Take out the sections and rinse them with deionized water to ensure no residual deparaffinization antigen retrieval solution is left (Do not rinse directly at the tissue to avoid tissue damage).
(5) Perform subsequent experiments with reference to the standard IHC experimental procedures.
Precautions
1. The optimal immersion time of sections in the deparaffinization antigen retrieval working solution needs to be optimized by the user according to different samples.
2. Operators must receive professional training and read this instruction manual carefully before use.
3. This product is exclusively for deparaffinization and antigen retrieval in immunohistochemistry and no other uses are permitted.
4. The deparaffinization and antigen retrieval methods described in this manual are standard protocols. Users may adjust the protocols or perform pre-experiments according to the specific properties of sections and types of antigens. Insufficient deparaffinization or antigen retrieval may lead to uneven staining or reduced staining intensity.
5. Over-retrieval may cause tissue damage or detachment.
6. The solution must be cooled naturally after heating; otherwise, the antigen retrieval effect will be impaired.
7. For your safety and health, wear a lab coat and disposable gloves during operation.
8. Use the reagent as soon as possible after opening to avoid compromising the results of subsequent experiments.
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