产品介绍
1. 外观:黄色液体
2. 纯度:≥99% (HPLC) 超纯级,MW=C16H17Cl2N5=350.25
3. 产品参数:Ex(nm) 358 Em(nm) 461 Solvent: Water
产品描述
DAPI是一种可对DNA 染色的细胞核染色试剂,它在嵌入双链DNA后释放蓝色荧光。DAPI常用于细胞凋亡检测,染色后用荧光显微镜观察或流式细胞仪检测。DAPI也常用于普通的细胞核染色以及某些特定情况下得双链DNA染色。尽管DAPI不能通过活细胞膜,但却能穿透扰乱的细胞膜而对核染色。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-DNA复合物的激发和发射波长分别为360nm和460nm。
本DAPI染色液可以直接用于固定细胞或组织的细胞核染色。
使用方法
(1)取适量1mM DAPI水溶液加到PBS*中,制备成10-50 µM的DAPI溶液。
推荐以下PBS的配制方法:NaCl: 8.00g KCl: 0.20g Na2HPO4 .12H2O: 2.9g KH2PO4: 0.2g 以上试剂溶解于1000 ml纯水中。
(2)将1/10培养基体积的DAPI溶液加入到细胞培养基中。(也可以用1/10浓度的DAPI缓冲液代替培养基。)
(3)在37℃培养细胞10-20分钟。
(4)用PBS或合适的缓冲液洗细胞两次。
(5)用带有360 nm激发波长,460 nm发射波长的滤光片的荧光显微镜观察细胞。
储存条件:-20℃避光保存,有效期一年。
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。
Product introduction
1. Appearance: yellow liquid
2. Purity: ≥99% (HPLC) ultra-pure grade, MW=C16H17Cl2N5=350.25
3. Product parameters: Ex(nm) 358 Em(nm) 461 Solvent: Water
Product description
DAPI is a nuclear staining reagent that can stain DNA. It releases blue fluorescence after being inserted into double-stranded DNA. DAPI is often used for cell apoptosis detection. After staining, it is observed with a fluorescence microscope or detected by flow cytometry. DAPI is also commonly used for general nuclear staining and double-stranded DNA staining in some specific cases. Although DAPI cannot pass through living cell membranes, it can penetrate disturbed cell membranes to stain nuclei. DAPI has a high photobleaching tolerance level and can be used to detect yeast mitochondrial DNA, chloroplast DNA, virus DNA, microplasm DNA and chromosomal DNA. The excitation and emission wavelengths of the DAPI-DNA complex are 360nm and 460nm, respectively.
The DAPI staining solution can be directly used for staining the nucleus of fixed cells or tissues.
Instructions
(1) Take an appropriate amount of 1mM DAPI aqueous solution and add it to PBS* to prepare a 10-50 µM DAPI solution.
The following PBS preparation method is recommended: NaCl: 8.00g KCl: 0.20g Na2HPO4 .12H2O: 2.9g KH2PO4: 0.2g The above reagents are dissolved in 1000 ml of pure water.
(2) Add 1/10 of the volume of the DAPI solution to the cell culture medium. (You can also use 1/10 concentration of DAPI buffer instead of the medium.)
(3) Incubate the cells at 37°C for 10-20 minutes.
(4) Wash the cells twice with PBS or a suitable buffer.
(5) Observe the cells with a fluorescence microscope with a filter with excitation wavelength of 360 nm and emission wavelength of 460 nm.
Storage conditions: Store at -20°C and protected from light, valid for one year. …
Precautions
1) DAPI is irritating to the human body, please pay attention to proper protection.
2) Fluorescent dyes all have the problem of quenching. It is recommended to complete the test on the same day after dyeing.
3) To slow down fluorescence quenching, anti-fluorescence quenching mounting solution can be used.
4) For your safety and health, please wear lab coats and disposable gloves for operation.