组织在制作过程中由于化学试剂的作用封闭了抗原，又由于热作用致使部分抗原的肽链发生扭曲，致使在免疫组化的染色过程中不能将其显示出来，为了解决上述的问题，利用化学试剂和热作用将这些抗原重新暴露出来或修正过来的过程称为抗原修复。柠檬酸盐、EDTA、Tris等缓冲液在热的条件下可以使被福尔马林固定液屏蔽的抗原重新暴露出来，同时又不会对抗原表位造成破坏，从而提高抗原的检出率，降低背景染色，提高诊断的准确率。
抗原修复有多种方法，主要方法可简要归纳为加热修复和非加热抗原修复两大类。非加热抗原修复方法包括酶消化、真空负压、酸水解等方法。目前主要是酶消化法，酶消化是以化学的方法来打断醛键，修复抗原，在免疫组织与免疫组织化学中有时经福尔马林等醛基固定液过度固定的标本，常会产生过量的醛基遮盖抗原，影响一抗与抗原的结合，用蛋白酶溶液消化，可以起到暴露抗原的作用，因而在实验室中胃蛋白酶消化液常被用于组织切片的抗原修复。该试剂仅用于科研领域，不适用于临床诊断或其他用途。
自备材料：
1、蒸馏水、PBS磷酸盐缓冲液 (0.01 mol/L, pH 7.2-7.4)
2、恒温箱
操作步骤 (仅供参考)：
1、切片经二甲苯或脱蜡透明液脱蜡至水。
2、切片入H₂O₂甲醇溶液处理切片10 min。
3、自来水洗，蒸馏水洗。
4、PBS洗3次，每次1 min。
5、滴加或将玻片浸入胃蛋白酶抗原修复液，孵育20 min。
6、PBS洗3次，每次2 min。
7、按选好的免疫组化染色方法进行染色。
注意事项：
1、由于组织固定方法和时间长短与强度不同，抗原修复所需的消化时间会有很大差异，实验人员应依据免疫染色的结果相应的调整，确定最佳消化时间。
2、胃蛋白酶抗原修复液属蛋白酶制剂，室温放置过久易导致酶活性的降低或丧失，反复冻融亦会导致酶活性的丧失。
3、本品不含防腐剂，一经开启不宜4 ℃长期保存。
4、为了您的安全和健康，请穿实验服并戴一次性手套操作。
5、试剂开封后请尽快使用，以防影响后续实验效果。
During tissue processing, antigens are masked by chemical reagents, and thermal treatment can cause distortion of the peptide chains of some antigens, making them undetectable in immunohistochemical staining. To solve these problems, the process of re‑exposing or restoring these antigens using chemical reagents and heat is called antigen retrieval. Buffers such as citrate, EDTA, and Tris, under heated conditions, can re‑expose antigens masked by formalin fixative without damaging antigenic epitopes. This improves antigen detection rate, reduces background staining, and enhances diagnostic accuracy.
There are multiple methods of antigen retrieval, which can be briefly divided into two main categories: heat‑induced epitope retrieval and non‑heating antigen retrieval. Non‑heating methods include enzymatic digestion, vacuum‑assisted retrieval, acid hydrolysis, etc. Currently, enzymatic digestion is the most commonly used. Enzymatic digestion uses chemical methods to break aldehyde bonds and retrieve antigens. In immunohistology and immunohistochemistry, specimens over‑fixed by aldehyde fixatives such as formalin often show excessive aldehyde masking of antigens, which hinders the binding of primary antibodies to antigens. Digestion with protease solution can expose the antigens. Therefore, pepsin solution is commonly used in laboratories for antigen retrieval of tissue sections. This product is for research use only and is not suitable for clinical diagnosis or other applications.
Materials Provided by User
1. Distilled water, PBS phosphate‑buffered saline (0.01 mol/L, pH 7.2-7.4)
2. Constant temperature incubator
Protocol (For Reference Only)
1. Deparaffinize sections with xylene or deparaffinizing solution, then hydrate to water.
2. Incubate sections in H₂O₂ methanol solution for 10 min.
3. Rinse with tap water, then distilled water.
4. Wash with PBS 3 times, 1 min each.
5. Add pepsin antigen retrieval solution onto sections or immerse slides in the solution; incubate for 20 min.
6. Wash with PBS 3 times, 2 min each.
7. Stain according to the selected immunohistochemical staining method.
Precautions
1. Digestion time for antigen retrieval varies greatly depending on tissue fixation method, duration, and intensity. Researchers should adjust accordingly based on staining results to determine the optimal digestion time.
2. Pepsin antigen retrieval solution is a protease preparation. Prolonged storage at room temperature may reduce or abolish enzyme activity; repeated freeze‑thaw cycles also cause loss of activity.
3. This product contains no preservatives. Once opened, it is not suitable for long‑term storage at 4 ℃.
4. For your safety and health, please wear a lab coat and disposable gloves during operation.
5. Please use the reagent promptly after opening to avoid affecting subsequent experimental results.
ZJ26F0434103
危险品化学品经营许可证（带存储）