|
| 產(chǎn)地 | 北京 |
| 品牌 | Solarbio |
| 貨號 | D8200 |
| 用途 | DNA鏈熒光染色 |
| 英文名稱 | DAPI，4’,6-Diamidino-2-phenylindole dihydrochloride (DAPI) |
| 包裝規(guī)格 | 10mg |
| 純度 | HPLC≥90%% |
| CAS編號 | 28718-90-3 |
| 保質(zhì)期 | 3年 |
| 是否進口 | 否 |
| 有效期 | 3年 |
|---|---|
| 溶解性 | 1 mg/mL in water |
| 別名 | 2-(4-Amidinophenyl)-6-indolecarbamidine dihydrochloride |
| 英文名稱 | DAPI，4’,6-Diamidino-2-phenylindole dihydrochloride (DAPI) |
| CAS | 28718-90-3 |
| 分子式 | C16H15N5·2HCl |
| 分子量 | 350.25 |
| 儲存條件 | 2-8℃ |
| 危險代碼 | R：36/37/38 S：26-36 |
| 純度 | HPLC≥90% |
| 外觀（性狀） | 黃色粉末 |
| 單位 | 支 |
DNA鏈熒光染色。
DAPI是一種可以穿透細胞膜的藍色熒光染料。和雙鏈DNA結(jié)合后可以產(chǎn)生比DAPI自身強20多倍的熒光。和EB(ethidium bromide)相比，對雙鏈DNA的染色靈敏度要高很多倍。
DAPI染色常用于細胞凋亡檢測，染色后用熒光顯微鏡觀察或流式細胞儀檢測。DAPI也常用于普通的細胞核染色以及某些特定情況下的雙鏈DNA染色。
DAPI的*激發(fā)波長為340nm，*發(fā)射波長為488nm；DAPI和雙鏈DNA結(jié)合后，*激發(fā)波長為364nm，*發(fā)射波長為454nm。
本DAPI溶液用水配制，加熱有助于溶解。
用于細胞核染色時，推薦的DAPI工作濃度為0.5-10μg/ml。
運輸與保存方法
常溫運輸。粉末在室溫下穩(wěn)定，需避光儲存。
注意事項
1)DAPI對人體有一定刺激性，請注意適當防護。
2)熒光染料都存在淬滅的問題，建議染色后盡量當天完成檢測。
3)為減緩熒光淬滅可以使用抗熒光淬滅封片液。
4)為了您的安全和健康，請穿實驗服并戴一次性手套操作。
配制母液
用雙蒸水溶解，終濃度為1mg/ml。本產(chǎn)品不可以用緩沖液直接溶解。-20℃可保存1年，建議分裝保存，避免反復凍融。
使用方法
1.配制工作液：用雙蒸水或PBS稀釋母液，配制成所需要的工作的濃度（0.5-10μg/ml）。
2.固定的細胞或組織染色：
對于固定的細胞或組織樣品，固定后，適當洗滌去除固定劑。DAPI染色通常在其他染色的*進行。如果不需要進行其它染色，則直接進行DAPI 染色。
a)對于貼壁細胞或組織切片：加入適量DAPI染色液，覆蓋住樣品即可。
對于懸浮細胞：至少加入待測染色樣品體積3倍的染色液，混勻。室溫放置3-5分鐘。
b)吸除DAPI染色液，用TBST、PBS或生理鹽水洗滌2-3次，每次3-5分鐘。
c)直接在熒光顯微鏡下觀察或封片后熒光顯微鏡下觀察。激發(fā)波長360nm，發(fā)射波長460nm。
3. 活細胞或組織染色：
a)細胞培養(yǎng)物中加入適量DAPI染色液，約1/10細胞培養(yǎng)基體積，必須充分覆蓋住待染色的樣品。通常對于六孔板一個孔需加入1ml染色液，對于96孔板一個孔需加入100μl染色液。
b)在 37℃培養(yǎng)細胞 10～20 分鐘。
c)用 PBS或合適的緩沖液洗細胞兩次。
d)直接在熒光顯微鏡下觀察或封片后熒光顯微鏡下觀察。激發(fā)波長360nm，發(fā)射波長460nm。
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