| 產(chǎn)品編號(hào) | IHC0101M | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| 英文名稱 | Mouse GFAP Ready-To-Use IHC Kit | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| 中文名稱 | 小鼠膠質(zhì)纖維酸性蛋白即用型免疫組化試劑盒 | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| 別 名 | Astrocyte Marker; FLJ45472; GFAP; Glial Fibrillary Acidic Protein; Intermediate filament protein; GFAP_HUMAN. | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| 種 屬 | mouse | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| 應(yīng)用范圍 | Immunohistochemistry | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| 適用樣品基質(zhì) | FFPE tissue | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| 描述 | IHC0101 is a ready-to-use IHC kit for staining of GFAP. The kit provides all reagents, from antigen retrieval to cover slip mounting, that require little to no diluting or handling prior to use. Simply apply the reagents to your sample slide according to the protocol and you're steps away from obtaining high-quality IHC data. | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| 保存條件 | Please store components at the temperatures indicated on the individual tube labels. The kit is stable for 6 months from the date of receipt. | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| 注意事項(xiàng) | This product as supplied is intended for research use only, not for use in human, therapeutic or diagnostic applications. | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| 產(chǎn)品介紹 |
產(chǎn)品組分及規(guī)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.
石蠟包埋組織的免疫組織化學(xué)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(fù)3次。 2. 抗原修復(fù) 沸水浴修復(fù)：將100×抗原修復(fù)緩沖液（試劑2）用蒸餾水稀釋成1×抗原修復(fù)緩沖液，放入修復(fù)盒中并提前加熱至95-100℃（注意蓋好以防液體蒸發(fā)），然后將切片放入修復(fù)盒中，在沸水浴環(huán)境中保持外沸狀態(tài)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(fù)3次。 3. 阻斷內(nèi)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(nèi)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(fù)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(nèi)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滴小鼠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(fù)溫 4℃孵育過夜后，室溫下復(fù)溫15 min (若在室溫下孵育一抗，則直接進(jìn)入下一步清洗)；用PBS洗滌5 min，重復(fù)3次。 7. 二抗孵育 用吸水紙吸玻片上多余的液體，加入2-4滴HRP標(biāo)記羊抗兔IgG工作液（試劑6），置于濕盒中，37℃孵育1-2 h；用PBS洗滌5 min，重復(fù)3次。 8. 顯色 用吸水紙吸去玻片上多余的液體，在每張切片上滴加約50 μL新配制的DAB工作液（試劑7:試劑8:PBS=1:1:18），作用3-5 min。顯微鏡下觀察結(jié)果，達(dá)到合適的顯色強(qiáng)度后，用蒸餾水沖洗切片以終止反應(yīng)，用蒸餾水沖洗5 min，重復(fù)3次。 9. 復(fù)染 滴加適量復(fù)染試劑（試劑9）復(fù)染3-5 min，蒸餾水沖洗5 min，滴加鹽酸酒精分化約30 s，蒸餾水洗滌5 min，重復(fù)2次。 10. 脫水封片 將玻片依次置于不同濃度（70%、80%、90%、95%、100%）乙醇，各5 min；然后置于新鮮二甲苯中浸泡脫蠟3次，每次15 min。用吸水紙吸去多余的二甲苯，滴加適量封片劑（試劑10）在組織上，將蓋玻片蓋在組織上，避免產(chǎn)生氣泡。
注意事項(xiàng) 1. 建議檢測(cè)時(shí)進(jìn)行陰性及陽性對(duì)照，以提高實(shí)驗(yàn)的可靠性。 2. 本品中的配套試劑，請(qǐng)不要用其他生產(chǎn)商產(chǎn)品替換使用。 3. DAB為致癌物質(zhì)，請(qǐng)采取必要的防范措施。 4. PBS洗滌液（試劑1）配制后在4℃可保存一周；抗原修復(fù)液（試劑2）及顯色劑（試劑7和8）的工作液需每次實(shí)驗(yàn)時(shí)現(xiàn)用現(xiàn)配。 *5. 發(fā)表論文時(shí)引用本產(chǎn)品的寫作建議 "IHC0101M, Bioss Antibodies"。引用示例: “Mouse tissue sections using Mouse GFAP IHC Kit (IHC0101M, Bioss Antibodies) were stained for GFAP according to the manufacturer's instructions.” |
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| 產(chǎn)品圖片 |
Immunohistochemical analysis of paraffin embedded mouse brain tissue slide using IHC0101M (Mouse GFAP IHC Kit).
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