产品介绍：
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。
Tris-EDTA抗原修复液(10×，pH9.0)可以有效去除醛类固定试剂导致的蛋白之间的交联，充分暴露石蜡切片等样品中的抗原表位，可以用于石蜡切片、冰冻切片等样品使用多聚甲醛、甲醛或其它醛类试剂固定后的抗原修复。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进，可考虑进行抗原修复。按照每个片子需要10ml抗原修复液(1×)计算，100ml抗原修复液(10×)可以用于100个样本的抗原修复。
自备材料：
1、 系列乙醇
2、 双蒸水或去离子水
3、 加热设备
4、 免疫染色洗涤液
使用方法：
（一） 石蜡切片：
1、 脱蜡至水
①二甲苯3次，每次3~5min。
②无水乙醇脱水2次，每次3~5min。
③95%的乙醇，3~5min。
④90%的乙醇，3~5min。
⑤80%的乙醇，3~5min。
⑥70%的乙醇，3~5min。
⑦蒸馏水冲洗2次，每次3~5min。
2、 抗原修复
①用去离子水或双蒸水稀释Tris-EDTA抗原修复液(10×,pH9.0)至1×。
②将切片浸泡在Tris抗原修复液（1×）中，95℃或沸水加热约10~30min。
③抗原修复液（1×）用前需预热到95~100℃。如果使用微波炉加热，避免暴沸和过多的水分蒸发。随后大约在20~30min内冷却至室温。
3、 免疫染色洗涤液洗涤1~2次，每次3~5min。
4、 封闭等后续的免疫染色步骤。
（二） 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、 浸泡在抗原修复液（1×）中，最佳的加热时间需根据不同的样品和目的蛋白自行摸索。
2、 为了您的安全和健康，请穿好实验服并佩戴一次性手套和口罩操作。
3、 本产品仅限于专业人员的科学研究用，不得用于临床诊断或治疗，不得用于食品或药品，不得存放于普通住宅内。
注意:
1.本产品仅供科研使用。请勿用于医药、临床诊断或治疗。食品及化妆品等用途。请勿存放于普通住宅区。
2.为了您的安全和健康，请穿好实验服并佩戴一次性手套和口罩操作。
3.实验结果可由多种因素影响，相关处理只限于产品本身，不涉及其他赔偿。
参考文献 (1)
We previously showed that the rupture of Bowman’s capsule (BC) promotes the progression of crescentic glomerulonephritis by enhancing the entry of CD8+ T cells into the glomeruli. In the present study, we utilized digital spatial profiling to simultaneously profile the altered abundances of the messenger RNA (mRNA) transcripts and proteins in the glomerular and periglomerular areas of four biopsy samples of anti-neutrophil cytoplasmic autoantibody-associated glomerulonephritis (ANCA-GN) and two biopsy specimens of minimal change disease (MCD) controls. The paraffin-embedded biopsy samples were stained with collagen IV, CD45, and SYTO 13 to distinguish the glomeruli with periglomerular infiltration but intact BC, with focal BC rupture, and with extensive rupture of BC and glomeruli without crescent formation and leukocytic infiltration in ANCA-GN. By assessing multiple discrete glomerular areas, we found that the transcript expression levels of the secreted phosphoprotein-1 and its receptor CD44 were upregulated significantly in the glomeruli with more severe ruptures of BC, and their expression levels correlated positively with the fibrotic markers. We also found that both alternative and classic complement pathways were activated in the glomeruli from patients with ANCA-GN. Furthermore, M1 macrophages were involved mostly in the early stage of BC rupture, while M2 macrophages were involved in the late stage and may contribute to the fibrosis process of the crescents. Finally, loss of glomerular cells in ANCA-GN was likely mediated by apoptosis. Our results show that digital spatial profiling allows the comparative analysis of the mRNA and protein profiles in individual glomeruli affected differently by the disease process and the identification of potential novel mechanisms in ANCA-GN.
