產品概述:
產品參數
外觀:黃色粉末
Ex/Em(結合DNA)=360/460 nm
CAS號:28718-90-3
分子量:350.3
分子式:C16H17Cl2N5
分子結構圖:
儲存條件
-20℃避光保存,有效期見外包裝。
產品介紹
以貼壁細胞(96孔板)舉例,每孔100 μL染色工作液,染色工作液濃度按5 μg/mL計算,10 mg配置為工作液大概可以用于20000個孔的染色。
注意事項
1. 使用前請將產品瞬時離心至管底,再進行后續實驗。
2. 相較于染色細菌,DAPI染料對哺乳動物細胞染色更加靈敏,染色死活細菌時推薦在PBS或150 mM NaCl中溶解為終濃度10 μg/mL的染色液,室溫染色30 min。通常對于死細胞的染色要比活細胞染色亮度高。
3. 用于細胞核染色時,推薦DAPI工作濃度為0.5-10 μg/mL。
4. DAPI對人體有害,請穿實驗服并戴一次性手套操作,注意適當防護。
5. 熒光染料均存在淬滅問題,請盡量注意避光,以減緩淬滅。
常見問題解答:
◆DAPI和Hoechst 染料非常相似,該如何選用?
DAPI是一種非常普通的藍色核復染熒光染料,能對固定和通透的細胞和組織的細胞核進行非常明亮的標記。遺憾的是,人們普遍認為它是介于半通透性到非通透性之間的染色劑,對活細胞的染色效果也不一致。Hoechst 33342染料是細胞通透性染料,與DAPI染色有相似的染色結合機制和熒光顏色;它是活細胞成像的首選,且對固定細胞的標記效果和DAPI一樣好。
◆DAPI染色的工作液濃度一般為多少,要怎么稀釋?
該試劑用于細胞染色時,需要根據具體的實驗目的確定工作液濃度,常用的濃度0.5-10μg/ml,直接用水稀釋即可。
◆在細胞膜和細胞核雙染實驗中,細胞膜染色用DiI,細胞核染色用DAPI,細胞固定后,先染色DiI還是先染色DAPI?
先用DiI對細胞膜進行染色,后用DAPI染核。
◆DAPI和Hoechst可以用來示蹤細胞嗎?
不可以,這些染料與DNA和RNA的結合會影響核酸的正常功能,擾亂轉錄和增殖。
◆DAPI能否用來染色植物細胞的原生質體?
可以,有文獻報道使用DAPI染色原生質體,但是原生質體比較脆弱,DAPI對原生質體的殺傷力可能比較大。
◆該試劑可對酵母菌進行熒光標記嗎?
有文獻用DAPI染酵母樣品,但我們目前沒有相關經驗,建議查看相關文獻資料進行參考。
◆該試劑可以染活細胞嗎?
DAPI染色時,細胞均需要固定操作,對于活細胞的染色效果較差。
◆請問可以用于石蠟切片的免疫組化,進行染核嗎?
可以的。
◆DAPI在配置成溶液后可以穩定存放多久?
儲液建議用雙蒸水配置成1-5mg/ml,若經常使用建議分裝成小規格,避免反復凍融,-20℃避光保存,可穩定存放1年;染色工作液建議用PBS稀釋成0.5-10ug/ml, 4℃避光保存,可穩定存放6個月。
使用本產品的文獻:
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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)
應用方向:復染
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)
應用方向:復染
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
應用方向:復染
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
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cellular & molecular immunology
7.DHA alleviates diet-induced skeletal muscle fber remodeling via FTO/m6A/DDIT4/PGC1α signaling
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參考文獻:
1.Assay for nanogram quantities of DNA in cellular homogenates
應用方向:DNA含量測定2.A simple cytochemical techniqUE for demonstration of DNA in cells infected with mycoplasmas and viruses
應用方向:診斷細胞培養的支原體感染
應用方向:流式細胞術中細胞核含量測定及染色體分選
4.Loss of HSulf-1 up-regulates heparin-binding growth factor signaling in cancer
應用方向:評估細胞凋亡
5.Direct measurement of femtogram amounts of DNA in cells and chloroplasts by quantitative microspectrofluorometry.
應用方向:免疫熒光法和原位雜交法檢測細胞核和細胞器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
應用方向:免疫熒光法和原位雜交法檢測細胞核和細胞器DNA
7.A simple cytochemical techniqUE for demonstration of DNA in cells infected with mycoplasmas and viruses
應用方向:取代溴化乙錠對瓊脂糖凝膠中的DNA進行染色
8.Comparison of ethidium bromide and 4', 6'-diamidino-2-phenylindole as quantitative fluorescent stains for DNA in agarose gels
應用方向:取代溴化乙錠對瓊脂糖凝膠中的DNA進行染色
9.Localization of polyphosphates in Saccharomyces fragilis, as revealed by 4′, 6-diamidino-2-phenylindole fluorescence
應用方向:DAPI可與多磷酸鹽和其他聚陰離子,硫酸葡聚糖和SDS結合
10.InflUEnce of S-adenosylmethionine on 4′, 6-diamidino-2-phenylindole dihydrochloride (DAPI)-induced fluorescence of polyphosphate in the yeast vacuole
應用方向:DAPI可與多磷酸鹽和其他聚陰離子,硫酸葡聚糖和SDS結合
11.Fluorescent Complexes of DNA With DAPI 4',6-diamidine-2-phenyl indole.2HCl or DCI 4',6-dicarboxyamide-2-phenyl Indole
應用方向:DAPI可與多磷酸鹽和其他聚陰離子,硫酸葡聚糖和SDS結合