产品说明书
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已发表文献
产品描述
DAPI，也称DAPI dihydrochloride，是一种常用的核酸染料，可以和双链DNA富含AT序列的小沟结合，产生比自身强20多倍的蓝色荧光。和EB(ethidium bromide)相比，DAPI对双链DNA的染色灵敏度要高很多倍。DAPI也可对RNA进行染色，染色机理是其可以选择性嵌入“AU序列”并发出荧光。相比DAPI-dsDNA(Ex/Em=358 nm/461 nm)，DAPI-RNA具有较长的最大发射波长(500 nm)，其荧光亮度仅有DAPI-dsDNA的20%。
尽管DAPI不能通过活细胞膜，但可以通过提高浓度使之进入活细胞。DAPI具有很高的光漂白承受水平，能用来检测酵母线粒体DNA，叶绿体DNA，病毒DNA，Microplasm DNA以及染色体DNA。
DAPI典型的蓝色荧光特性使其非常普遍的搭配其他绿色、黄色或红色荧光染料用于细胞生物学多色荧光标记技术，因此也常作为核酸和染色体的复染剂用于细胞凋亡检测、RNA原位杂交、直接或间接免疫检测等领域，其染色后可用荧光显微镜观察或流式细胞仪检测。推荐工作浓度为0.5-10 μg/mL。
产品性质
|
中文名称（Chinese Synonym） |
4’,6-联脒-2-苯基吲哚二盐酸盐 |
|
英文名称（English Synonym） |
2-(4-Amidinophenyl)-6-indolecarbamidine dihydrochloride; DAPI dihydrochloride 4’,6-Diamidino-2-phenylindole, dihydrochloride; |
|
分子式（Molecular Fomular） |
C16H15N5·2HCl |
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分子量（Molecular Weight） |
350.25 |
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CAS号（CAS NO.） |
28718-90-3 |
|
纯度（HPLC Purity） |
≥95% |
|
荧光光谱（Fluorescence Spectral） |
|
|
结构式（Structure） |
|
运输与保存方法
常温运输。粉末于4 ℃保存，需避光储存，有效期3年。
注意事项
1）DAPI对人体有一定刺激性，请注意适当防护。
2）荧光染料都存在淬灭的问题，建议染色后尽量当天完成检测。
3）为减缓荧光淬灭可以使用抗荧光淬灭封片液。
4）为了您的安全和健康，请穿实验服并戴一次性手套操作。
5）本产品仅作科研用途！
配制母液
用双蒸水溶解，终浓度为1-5 mg/mL。本产品不可以用缓冲液直接溶解。-20 ℃可保存1年，建议分装保存，避免反复冻融
使用方法
1. 配制工作液：用双蒸水或PBS稀释母液，配制成所需要的工作的浓度（0.5-10 μg/mL），工作液可于4 ℃保存6个月。
2. 固定的细胞或组织染色：对于固定的细胞或组织样品，固定后，适当洗涤去除固定剂。DAPI染色通常在其他染色的最后进行。如果不需要进行其它染色，则直接进行DAPI 染色。
a）对于贴壁细胞或组织切片：加入适量DAPI染色液，覆盖住样品即可。
对于悬浮细胞：至少加入待测染色样品体积3倍的染色液，混匀。室温放置3-5分钟。
b）吸除DAPI染色液，用TBST、PBS或生理盐水洗涤2-3次，每次3-5分钟。
c）直接在荧光显微镜下观察或封片后荧光显微镜下观察。激发波长360 nm，发射波长460 nm。
3. 活细胞或组织染色：
a）细胞培养物中加入适量DAPI染色液，约1/10细胞培养基体积，必须充分覆盖住待染色的样品。通常对于六孔板一个孔需加入1 mL染色液，对于96孔板一个孔需加入100 μL染色液。
b）在 37 ℃培养细胞 10～20 分钟。
c）用 PBS或合适的缓冲液洗细胞两次。
d）直接在荧光显微镜下观察或封片后荧光显微镜下观察。激发波长360 nm，发射波长460 nm。
相关产品
|
产品货号 |
产品编号 |
规格 |
|
40724ES10 |
10 g |
|
|
40729ES10 |
10 mg |
|
|
Hoechst 33258 染液（1 mg/mL） Hoechst 33258 Stain Solution (1 mg/mL) |
40730ES03 |
1 mL |
|
40731ES10 |
10 mg |
|
|
Hoechst 33342 染液（1mg/mL）Hoechst 33342 Stain Solution (1 mg/mL) |
40732ES03 |
1 mL |
|
40755ES64 |
150 T |
|
|
40710ES03 |
1 mL |
|
|
40711ES10 |
10 mg |
|
|
40722ES03 |
1 mg |
|
|
40745ES64 |
150 T |
|
|
36307ES08 |
5 mL |
|
|
36308ES11 |
4 mL |
|
|
40728ES03 |
1 mg (200 μL) |
HB190805
Q:说明书上写的加入 1/10 细胞培养基体积DAPI 是指 96 孔板中加入培养基体积的 1/10 吗？
A:染色时可先弃培养基，直接加入 100ul 染液，并且这产品不推荐染活细胞，那如果是染活细胞，建议用hoechest 染料。
Q．DAPI和Hoechst染料相当类似，如何二选一？
A：DAPI是非常通用的蓝色荧光染料，用于细胞核复染。对固定和透化的细胞/组织，具有非常明亮的细胞核标记。然而，此染料被认为是半渗透至不渗透的染料，用于活细胞染色的结合不一致。Hoechst 33342是一种细胞膜渗透性的染料，具有同DAPI的结合机制和荧光特征。非常适合用于活细胞成像，效果就如同DAPI用于固定细胞染色。
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