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產品組分及規格

背景

GFAP (Glial fibrillary acidic protein) is a member of the class III intermediate filament protein family. GFAP is heavily and specifically expressed in astrocytes and certain astroglia of the central nervous system, in satellite cells of peripheral ganglia, and in non-myelinating Schwann cells of peripheral nerves. In addition, neural stem cells strongly express GFAP. Antibodies to GFAP are very useful as markers of astrocytic cells. In addition, many types of brain tumor, presumably derived from astrocytic cells, heavily express GFAP. GFAP is also found in the lens epithelium, Kupffer cells of the liver, in some cells in salivary tumors and has been reported in erythrocytes. GFAP is used as a marker to distinguish astrocytes from other glial cells during development. Mutations in this gene cause Alexander disease, a rare disorder of astrocytes in the central nervous system. Alternative splicing of the GFAP gene results in multiple transcript variants encoding distinct isoforms.

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石蠟包埋組織的免疫組織化學方案

1.?脫蠟水化

石蠟切片置于新鮮二甲苯中浸泡脫蠟3次，每次15 min；依次置于不同濃度（100%、95%、90%、80%、70%）乙醇浸泡各5 min，再置于蒸餾水洗滌5 min，重復3次。

2.?抗原修復

沸水浴修復：將100×抗原修復緩沖液（試劑2）用蒸餾水稀釋成1×抗原修復緩沖液，放入修復盒中并提前加熱至95-100℃（注意蓋好以防液體蒸發），然后將切片放入修復盒中，在沸水浴環境中保持外沸狀態15 min，室溫自然冷卻；用PBS緩沖液（試劑1，將干粉溶解在2L蒸餾水中）清洗5 min，重復3次。

3.?阻斷內源性過氧化物酶

用吸水紙吸去玻片上多余的液體，用免疫組化筆在組織周圍畫圈，加入2-4滴內源性過氧化物酶阻斷劑（試劑3），室溫下置于濕盒中孵育15 min，用PBS洗滌5 min，重復3次。

4.?血清封閉

用吸水紙吸去玻片上多余的液體，加入2-4滴封閉工作液（試劑4），置于濕盒內37℃封閉20 min，以減少非特異性染色。

5.?一抗孵育

用吸水紙吸去玻片上多余的液體，加入2-4滴大鼠GFAP兔多抗工作液（試劑5），置于濕盒中，4℃孵育過夜或37℃孵育1-2 h。

6.?復