石蠟切片免疫組化步驟及所需生化試劑列表

2017-05-11

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下面詳細(xì)介紹實(shí)驗(yàn)步驟:

  第一步:石蠟切片置于67℃烘箱中,烘片2小時(shí),脫蠟至水,用pH7.4的PBS沖洗三次,每次3分鐘(3×3’)。

  第二步:取一定量pH=6.0檸檬酸鹽緩沖液,加入微波盒中,微波加熱至沸騰,將脫蠟水化后的組織切片置于耐高溫塑料切片架上,放入已沸騰的緩沖液中,中檔微波處理10分鐘,取出微波盒流水自然泠卻,從緩沖液中取出玻片,先用蒸餾水沖洗兩次,之后用PBS沖洗2×3’。(注:不是所有的抗體都需要微波修復(fù)的,視具體情況而定)

  第三步:每張切片加1滴3%H2O2,室溫下孵育10分鐘,以阻斷內(nèi)源性過氧化物酶的活性。PBS沖洗3×3’。

  在此免疫組化實(shí)驗(yàn)中可能需要用的試劑及儀器耗材包括:PBS、檸檬酸鹽緩沖液、蒸餾水、增強(qiáng)劑、酶標(biāo)抗鼠、兔聚合物、蘇木素、酒精、二甲苯、樹膠、鹽酸、二氨基聯(lián)苯胺。


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