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DAPI染液(5mg/ml)DAPI Stain Solution(5mg/ml)

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產(chǎn)品描述

DAPI，也稱DAPI dihydrochloride，是一種常用的核酸染料，可以和雙鏈DNA富含AT序列的小溝結(jié)合，產(chǎn)生比自身強(qiáng)20多倍的藍(lán)色熒光。和EB(ethidium bromide)相比，DAPI對(duì)雙鏈DNA的染色靈敏度要高很多倍。DAPI也可對(duì)RNA進(jìn)行染色，染色機(jī)理是其可以選擇性嵌入“AU序列”并發(fā)出熒光。相比DAPI-dsDNA(Ex/Em=358 nm/461 nm)，DAPI-RNA具有較長的最大發(fā)射波長(500 nm)，其熒光亮度僅有DAPI-dsDNA的20%。

盡管DAPI不能通過活細(xì)胞膜，但可以通過提高濃度使之進(jìn)入活細(xì)胞。DAPI具有很高的光漂白承受水平，能用來檢測酵母線粒體DNA，葉綠體DNA，病毒DNA，Microplasm DNA以及染色體DNA。

DAPI典型的藍(lán)色熒光特性使其非常普遍的搭配其他綠色、黃色或紅色熒光染料用于細(xì)胞生物學(xué)多色熒光標(biāo)記技術(shù)，因此也常作為核酸和染色體的復(fù)染劑用于細(xì)胞凋亡檢測、RNA原位雜交、直接或間接免疫檢測等領(lǐng)域，其染色后可用熒光顯微鏡觀察或流式細(xì)胞儀檢測。

本產(chǎn)品為溶液，濃度為5 mg/mL。另外提供粉末形式的DAPI（Cat No. 40727ES10）供客戶選購。

產(chǎn)品性質(zhì)

中文名稱（Chinese Synonym）	4’,6-聯(lián)脒-2-苯基吲哚二鹽酸鹽
英文名稱（English Synonym）	4’,6-Diamidino-2-phenylindole, dihydrochloride
CAS號(hào)（CAS NO.）	28718-90-3
分子式（Molecular Fomular）	C₁₆H₁₇C_l2N₅·2HCl
分子量（Molecular Weight）	350.25
熒光光譜（Fluorescence Spectral）	DAPI的Ex/Em=340 nm/488 nm; DAPI-DNA的Ex/Em=358 nm/461 nm

運(yùn)輸與保存方法
冰袋運(yùn)輸，5 mg/mL的DAPI儲(chǔ)存液于-20ºC避光保存，1年有效。

注意事項(xiàng)

1）DAPI對(duì)人體有一定刺激性，請(qǐng)注意適當(dāng)防護(hù)。

2）熒光染料都存在淬滅的問題，建議染色后盡量當(dāng)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）低濃度的DAPI不容易穿透細(xì)胞膜。

5）為了您的安全和健康，請(qǐng)穿實(shí)驗(yàn)服并戴一次性手套操作。

6）本產(chǎn)品僅作科研用途！

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（0.5-10 μg/mL）。

2. 固定的細(xì)胞或組織染色：

對(duì)于固定的細(xì)胞或組織樣品，固定后，適當(dāng)洗滌去除固定劑。DAPI染色通常在其他染色的最后進(jìn)行。如果不需要進(jìn)行其它染色，則直接進(jìn)行DAPI 染色。

a) 對(duì)于貼壁細(xì)胞或組織切片：加入適量DAPI染色液，覆蓋住樣品即可。

b) 對(duì)于懸浮細(xì)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。

c) 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。

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360 nm，發(fā)射波長460 nm。

3. 活細(xì)胞或組織染色：

a) 細(xì)胞培養(yǎng)物中加入適量DAPI染色液，約1/10細(xì)胞培養(yǎng)基體積，必須充分覆蓋住待染色的樣品。通常對(duì)于六孔板一個(gè)孔需加入1 mL染色液，對(duì)于96孔板一個(gè)孔需加入100 μL染色液。

b) 在37℃培養(yǎng)細(xì)胞 10～20 分鐘。

c) 用 PBS或合適的緩沖液洗細(xì)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360 nm，發(fā)射波長460 nm。

相關(guān)產(chǎn)品

產(chǎn)品貨號(hào)	產(chǎn)品編號(hào)	規(guī)格
Trypan blue 臺(tái)盼藍(lán)	40724ES10	10 g
Hoechst 33258	40729ES10	10 mg
Hoechst 33258 染液（1 mg/mL） Hoechst 33258 Stain Solution (1 mg/mL)	40730ES03	1 mL
Hoechst 33342	40731ES10	10 mg
Hoechst 33342 染液（1 mg/mL）Hoechst 33342 Stain Solution (1 mg/mL)	40732ES03	1 mL
Propidium iodide Staining Solution（1mg/mL）碘化丙啶染液（1 mg/mL）	40710ES03	1 mL
PI (Propidium iodide) 碘化丙啶	40711ES10	10 mg
7-AAD 7-氨基放線菌素D	40722ES03	1 mg
7-AAD Viability Staining Solution 7-AAD細(xì)胞活力染色液	40745ES64	150 T
Fluoromount-GTM 抗熒光淬滅封片劑（水溶性）	36307ES08	5 mL
DAPI Fluoromount-GTM 抗熒光淬滅封片劑（含DAPI，水溶性）	36308ES11	4 mL
DAPI染液（5 mg/mL）	40728ES03	1 mg (200 μL)

HB200211

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