组织在制作过程中，由于化学试剂的作用封闭了抗原，又由于热的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，利用化学试剂和热的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或Tris等缓冲液在热的条件下可以使被福尔马林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。
在热诱导抗原修复法出现之前，胰蛋白酶、蛋白酶K、胃蛋白酶等酶类的诱导的表位修复是最常用的方法。胰蛋白酶抗原修复试剂盒能暴露石蜡切片等样品中的抗原表位，从而大大改善免疫染色效果，通常石蜡切片都需进行抗原修复处理，而冰冻切片可以不进行抗原修复处理，抗原修复可以提高石蜡切片的免疫染色效果，亦可以不同程度的提高冰冻切片的染色效果；当冰冻切片免疫染色效果不理想时，可考虑进行抗原修复。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、系列乙醇、双蒸水或去离子水、免疫染色洗涤液
2、加热设备
操作步骤 (仅供参考)：
(一) 石蜡切片
1、脱蜡至水
①二甲苯或脱蜡透明液脱蜡3次，每次3-5 min。
②无水乙醇脱水2次，每次3-5 min。
③95%的乙醇，3-5 min。
④90%的乙醇，3-5 min。
⑤80%的乙醇，3-5 min。
⑥70%的乙醇，3-5 min。
⑦蒸馏水冲洗2次，每次3-5 min。
2、抗原修复：将切片浸泡在胰蛋白酶溶液中，37 ℃孵育10-25 min。
3、TB漂洗液洗涤1-2次，自来水洗涤，每次3-5 min，彻底漂洗干净。
4、进行后续的免疫染色步骤。
(二) 冰冻切片
1、免疫染色洗涤液或自来水洗涤切片5 min。
2、抗原修复：将切片浸泡在胰蛋白酶溶液中，37 ℃孵育10-25 min。
3、TB漂洗液洗涤1-2次，自来水洗涤，每次3-5 min，彻底漂洗干净。
4、进行后续的免疫染色步骤。
(三) 其它样品
其它样品参考石蜡切片或冰冻切片进行操作。
注意事项：
1、胰蛋白酶溶液恢复至室温后再使用，但应避免反复冻融。
2、浸泡在胰蛋白酶溶液中，最佳孵育时间需根据不同的样品和目的蛋白自行摸索。
3、为了您的安全和健康，请穿实验服并戴一次性手套操作。
4、试剂开封后请尽快使用，以防影响后续实验效果。
During tissue preparation, antigens are masked by chemical reagents, and some antigenic peptide chains are distorted due to heat, making them undetectable in immunohistochemical staining. To solve these problems, the process of re‑exposing or correcting these antigens using chemical reagents and heat is called antigen retrieval. Buffers such as citrate, EDTA, or Tris can re‑expose formalin‑masked antigens under heating conditions without damaging antigenic epitopes, thereby improving antigen detection rate, reducing background staining, and enhancing diagnostic accuracy.
Before the advent of heat‑induced antigen retrieval, epitope retrieval induced by enzymes such as trypsin, proteinase K, and pepsin was the most commonly used method. The Trypsin Antigen Retrieval Kit can expose antigenic epitopes in samples such as paraffin sections, greatly improving immunostaining results. In general, paraffin sections require antigen retrieval, while frozen sections may not. Antigen retrieval can enhance immunostaining of paraffin sections and also improve staining of frozen sections to varying degrees. Antigen retrieval can be considered when immunostaining of frozen sections is unsatisfactory. This reagent is for research use only. Not for clinical diagnosis or other applications.
Materials Provided by User
1. Graded ethanol, double‑distilled water or deionized water, immunostaining washing solution
2. Heating equipment
Protocol (For Reference Only)
(I) Paraffin Sections
1. Deparaffinization and rehydration
① Deparaffinize with xylene or deparaffinization clearing solution 3 times, 3-5 min each.
② Dehydrate with absolute ethanol 2 times, 3-5 min each.
③ 95% ethanol, 3-5 min.
④ 90% ethanol, 3-5 min.
⑤ 80% ethanol, 3-5 min.
⑥ 70% ethanol, 3-5 min.
⑦ Rinse with distilled water 2 times, 3-5 min each.
2. Antigen retrieval: Incubate sections in Trypsin Solution at 37℃ for 10-25 min.
3. Wash 1-2 times with TB Washing Buffer and tap water, 3-5 min each, until thoroughly rinsed.
4. Proceed with subsequent immunostaining steps.
(II) Frozen Sections
1. Wash sections with immunostaining washing solution or tap water for 5 min.
2. Antigen retrieval: Incubate sections in Trypsin Solution at 37℃ for 10-25 min.
3. Wash 1-2 times with TB Washing Buffer and tap water, 3-5 min each, until thoroughly rinsed.
4. Proceed with subsequent immunostaining steps.
(III) Other Samples
Follow the procedure for paraffin or frozen sections as appropriate.
Precautions
1. Allow Trypsin Solution to equilibrate to room temperature before use; avoid repeated freeze‑thaw cycles.
2. The optimal incubation time in Trypsin Solution must be determined empirically for different samples and target proteins.
3. For your safety and health, wear a lab coat and disposable gloves during operation.
4. Use the reagent promptly after opening to avoid compromising experimental results.
ZJ26F0434102
危险品化学品经营许可证（带存储）