产品概述:
产品参数
外观:黄色粉末
Ex/Em(结合DNA)=360/460 nm
CAS号:28718-90-3
分子量:350.3
分子式:C16H17Cl2N5
分子结构图:
储存条件
-20℃避光保存,有效期见外包装。
产品介绍
以贴壁细胞(96孔板)举例,每孔100 μL染色工作液,染色工作液浓度按5 µg/mL计算,10 mg配置为工作液大概可以用于20000个孔的染色。
注意事项
1. 使用前请将产品瞬时离心至管底,再进行后续实验。
2. 相较于染色细菌,DAPI染料对哺乳动物细胞染色更加灵敏,染色死活细菌时推荐在PBS或150 mM NaCl中溶解为终浓度10 μg/mL的染色液,室温染色30 min。通常对于死细胞的染色要比活细胞染色亮度高。
3. 用于细胞核染色时,推荐DAPI工作浓度为0.5-10 μg/mL。
4. DAPI对人体有害,请穿实验服并戴一次性手套操作,注意适当防护。
5. 荧光染料均存在淬灭问题,请尽量注意避光,以减缓淬灭。
常见问题解答:
◆DAPI和Hoechst 染料非常相似,该如何选用?
DAPI是一种非常普通的蓝色核复染荧光染料,能对固定和通透的细胞和组织的细胞核进行非常明亮的标记。遗憾的是,人们普遍认为它是介于半通透性到非通透性之间的染色剂,对活细胞的染色效果也不一致。Hoechst 33342染料是细胞通透性染料,与DAPI染色有相似的染色结合机制和荧光颜色;它是活细胞成像的首选,且对固定细胞的标记效果和DAPI一样好。
◆DAPI染色的工作液浓度一般为多少,要怎么稀释?
该试剂用于细胞染色时,需要根据具体的实验目的确定工作液浓度,常用的浓度0.5-10μg/ml,直接用水稀释即可。
◆在细胞膜和细胞核双染实验中,细胞膜染色用DiI,细胞核染色用DAPI,细胞固定后,先染色DiI还是先染色DAPI?
先用DiI对细胞膜进行染色,后用DAPI染核。
◆DAPI和Hoechst可以用来示踪细胞吗?
不可以,这些染料与DNA和RNA的结合会影响核酸的正常功能,扰乱转录和增殖。
◆DAPI能否用来染色植物细胞的原生质体?
可以,有文献报道使用DAPI染色原生质体,但是原生质体比较脆弱,DAPI对原生质体的杀伤力可能比较大。
◆该试剂可对酵母菌进行荧光标记吗?
有文献用DAPI染酵母样品,但我们目前没有相关经验,建议查看相关文献资料进行参考。
◆该试剂可以染活细胞吗?
DAPI染色时,细胞均需要固定操作,对于活细胞的染色效果较差。
◆请问可以用于石蜡切片的免疫组化,进行染核吗?
可以的。
◆DAPI在配置成溶液后可以稳定存放多久?
储液建议用双蒸水配置成1-5mg/ml,若经常使用建议分装成小规格,避免反复冻融,-20℃避光保存,可稳定存放1年;染色工作液建议用PBS稀释成0.5-10ug/ml, 4℃避光保存,可稳定存放6个月。
使用本产品的文献:
1.Extracellular matrix stiffness controls osteogenic differentiation of mesenchymal stem cells mediated by integrin α5
Meiyu Sun, Guangfan Chi, Juanjuan Xu, Ye Tan, Jiayi Xu, Shuang Lv, Ziran Xu, Yuhan Xia, Lisha Li and Yulin Li
Stem Cell Research &Therapy (2018)
应用方向:复染
2.Transcriptomic and Quantitative Proteomic Analyses Provide Insights Into the Phagocytic Killing of Hemocytes in the Oyster Crassostrea gigas
Shuai Jiang, Limei Qiu, Lingling Wang, Zhihao Jia, Zhao Lv, Mengqiang Wang,Conghui Liu, Jiachao Xu and Linsheng Song
Frontiers in Immunology (2018)
应用方向:复染
3.Overcoming the Reticuloendothelial System Barrier to Drug Delivery with a “Don’t-Eat-Us” Strategy
Yixuan Tang,Xiaoyou Wang,Jie Li,Yu Nie,Guojian Liao,Yang Yu,Chong Li
ACS NANO
应用方向:复染
Qing Zhang, Jia Liu, Ruolan Li, Rong Zhao, Mengmeng Zhang, Shujun Wei, Dong Ran, Wei Jin, Chunjie Wu
Frontiers in Pharmacology
5.Antiepileptic Effects of Cicadae Periostracum on Mice and Its Antiapoptotic Effects in H2O2-Stimulated PC12 Cells via Regulation of PI3K/Akt/Nrf2 Signaling Pathways
Qing Zhang, Ruo-Lan Li, Ting Tao, Jia-Yi Sun, Jia Liu, Ting Zhang, Wei Peng, Chun-Jie Wu
Volume 2021
6.SARS-CoV-2 ORF10 suppresses the antiviral innate immune response by degrading MAVS through mitophagy
Xingyu Li, Peili Hou, Wenqing Ma, Xuefeng Wang, Hongmei Wang, Zhangping Yu, Huasong Chang, Tiecheng Wang, Song Jin, Xue Wang, Wenqi Wang, Yudong Zhao, Yong Zhao, Chunqing Xu, Xiaomei Ma, Yuwei Gao, Hongbin He
cellular & molecular immunology
7.DHA alleviates diet-induced skeletal muscle fber remodeling via FTO/m6A/DDIT4/PGC1α signaling
Wei Chen, Yushi Chen, Ruifan Wu, Guanqun Guo, Youhua Liu, Botao Zeng, Xing Liao, Yizhen Wang, Xinxia Wang
BMC Biology
8.Guanxin V alleviates acute myocardial infarction by restraining oxidative stress damage, apoptosis, and fibrosis through the TGF-β1 signalling pathway
Phytomedicine
参考文献:
1.Assay for nanogram quantities of DNA in cellular homogenates
应用方向:DNA含量测定2.A simple cytochemical techniqUE for demonstration of DNA in cells infected with mycoplasmas and viruses
应用方向:诊断细胞培养的支原体感染
应用方向:流式细胞术中细胞核含量测定及染色体分选
4.Loss of HSulf-1 up-regulates heparin-binding growth factor signaling in cancer
应用方向:评估细胞凋亡
5.Direct measurement of femtogram amounts of DNA in cells and chloroplasts by quantitative microspectrofluorometry.
应用方向:免疫荧光法和原位杂交法检测细胞核和细胞器DNA
6.SMAD2 and SMAD7 Involvement in the post-translational regulation of Muc4 via the transforming growth factor-β and interferon-γ pathways in rat mammary epithelial cells
应用方向:免疫荧光法和原位杂交法检测细胞核和细胞器DNA
7.A simple cytochemical techniqUE for demonstration of DNA in cells infected with mycoplasmas and viruses
应用方向:取代溴化乙锭对琼脂糖凝胶中的DNA进行染色
8.Comparison of ethidium bromide and 4', 6'-diamidino-2-phenylindole as quantitative fluorescent stains for DNA in agarose gels
应用方向:取代溴化乙锭对琼脂糖凝胶中的DNA进行染色
9.Localization of polyphosphates in Saccharomyces fragilis, as revealed by 4´, 6-diamidino-2-phenylindole fluorescence
应用方向:DAPI可与多磷酸盐和其他聚阴离子,硫酸葡聚糖和SDS结合
10.InflUEnce of S-adenosylmethionine on 4´, 6-diamidino-2-phenylindole dihydrochloride (DAPI)-induced fluorescence of polyphosphate in the yeast vacuole
应用方向:DAPI可与多磷酸盐和其他聚阴离子,硫酸葡聚糖和SDS结合
11.Fluorescent Complexes of DNA With DAPI 4',6-diamidine-2-phenyl indole.2HCl or DCI 4',6-dicarboxyamide-2-phenyl Indole
应用方向:DAPI可与多磷酸盐和其他聚阴离子,硫酸葡聚糖和SDS结合