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  • DAPI(4',6-二脒基-2-苯基吲哚二鹽酸鹽)

    產(chǎn)品貨號(hào): D4054

    產(chǎn)品規(guī)格: 10 mg

    目錄價(jià)(元):620

    推薦儀器:熒光顯微鏡

    大包裝詢價(jià)


產(chǎn)品概述:

產(chǎn)品參數(shù)
外觀:黃色粉末
Ex/Em(結(jié)合DNA)=360/460 nm
CAS號(hào):28718-90-3
分子量:350.3
分子式:C16H17Cl2N5
分子結(jié)構(gòu)圖:


儲(chǔ)存條件

-20℃避光保存,有效期見(jiàn)外包裝。

產(chǎn)品介紹

DAPI是一種可對(duì)DNA染色的細(xì)胞核染色試劑,它在嵌入雙鏈DNA后釋放藍(lán)色熒光,亮度增強(qiáng)約20倍。DAPI常用于細(xì)胞凋亡檢測(cè),染色后用熒光顯微鏡觀察或流式細(xì)胞儀檢測(cè)。DAPI具有很高的光漂白承受水平,能用來(lái)檢測(cè)酵母線粒體DNA、葉綠體DNA、病毒DNA以及染色體DNA等。在較低濃度(1 μg/mL)時(shí),DAPI不可滲透于活細(xì)胞,但可用作固定細(xì)胞或組織部分的核染色劑。在較高濃度(10 μg/mL)時(shí),DAPI可用于染色活細(xì)胞。
以貼壁細(xì)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(jì)算,10 mg配置為工作液大概可以用于20000個(gè)孔的染色。

注意事項(xiàng)
1. 使用前請(qǐng)將產(chǎn)品瞬時(shí)離心至管底,再進(jìn)行后續(xù)實(shí)驗(yàn)。
2. 相較于染色細(xì)菌,DAPI染料對(duì)哺乳動(dòng)物細(xì)胞染色更加靈敏,染色死活細(xì)菌時(shí)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(duì)于死細(xì)胞的染色要比活細(xì)胞染色亮度高。
3. 用于細(xì)胞核染色時(shí),推薦DAPI工作濃度為0.5-10 μg/mL。
4. DAPI對(duì)人體有害,請(qǐng)穿實(shí)驗(yàn)服并戴一次性手套操作,注意適當(dāng)防護(hù)。
5. 熒光染料均存在淬滅問(wèn)題,請(qǐng)盡量注意避光,以減緩淬滅。

說(shuō)明書(shū):


 D4054    

常見(jiàn)問(wèn)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(lán)色核復(fù)染熒光染料,能對(duì)固定和通透的細(xì)胞和組織的細(xì)胞核進(jìn)行非常明亮的標(biāo)記。遺憾的是,人們普遍認(rèn)為它是介于半通透性到非通透性之間的染色劑,對(duì)活細(xì)胞的染色效果也不一致。Hoechst 33342染料是細(xì)胞通透性染料,與DAPI染色有相似的染色結(jié)合機(jī)制和熒光顏色;它是活細(xì)胞成像的首選,且對(duì)固定細(xì)胞的標(biāo)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(xì)胞染色時(shí),需要根據(jù)具體的實(shí)驗(yàn)?zāi)康拇_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(xì)胞膜和細(xì)胞核雙染實(shí)驗(yàn)中,細(xì)胞膜染色用DiI,細(xì)胞核染色用DAPI,細(xì)胞固定后,先染色DiI還是先染色DAPI?
先用DiI對(duì)細(xì)胞膜進(jìn)行染色,后用DAPI染核。

DAPI和Hoechst可以用來(lái)示蹤細(xì)胞嗎?
不可以,這些染料與DNA和RNA的結(jié)合會(huì)影響核酸的正常功能,擾亂轉(zhuǎn)錄和增殖。

DAPI能否用來(lái)染色植物細(xì)胞的原生質(zhì)體?
可以,有文獻(xiàn)報(bào)道使用DAPI染色原生質(zhì)體,但是原生質(zhì)體比較脆弱,DAPI對(duì)原生質(zhì)體的殺傷力可能比較大。

該試劑可對(duì)酵母菌進(jìn)行熒光標(biāo)記嗎?
有文獻(xiàn)用DAPI染酵母樣品,但我們目前沒(méi)有相關(guān)經(jīng)驗(yàn),建議查看相關(guān)文獻(xiàn)資料進(jìn)行參考。

該試劑可以染活細(xì)胞嗎?
DAPI染色時(shí),細(xì)胞均需要固定操作,對(duì)于活細(xì)胞的染色效果較差。

請(qǐng)問(wèn)可以用于石蠟切片的免疫組化,進(jìn)行染核嗎?
可以的。


DAPI在配置成溶液后可以穩(wěn)定存放多久?

儲(chǔ)液建議用雙蒸水配置成1-5mg/ml,若經(jīng)常使用建議分裝成小規(guī)格,避免反復(fù)凍融,-20℃避光保存,可穩(wěn)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(wěn)定存放6個(gè)月。


使用本產(chǎn)品的文獻(xiàn):


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)
應(yīng)用方向:復(fù)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(yīng)用方向:復(fù)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(yīng)用方向:復(fù)染


4.A Network Pharmacology Approach to Investigate the Anticancer Mechanism and Potential Active Ingredients of Rheum palmatum L. Against Lung Cancer via Induction of Apoptosis
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 ZhangRuo-Lan LiTing TaoJia-Yi SunJia LiuTing ZhangWei PengChun-Jie Wu
Volume 2021

6
.SARS-CoV-2 ORF10 suppresses the antiviral innate immune response by degrading MAVS through mitophagy
Xingyu LiPeili HouWenqing MaXuefeng WangHongmei WangZhangping YuHuasong ChangTiecheng WangSong JinXue WangWenqi WangYudong ZhaoYong ZhaoChunqing XuXiaomei MaYuwei GaoHongbin 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
Bo Liang Xiao-Xiao Zhang Rui Li Yong-Chun Zhu Xiao-Jie Tian Ning Gu
Phytomedicine

參考文獻(xiàn):

1.Assay for nanogram quantities of DNA in cellular homogenates

應(yīng)用方向:DNA含量測(cè)定

2.A simple cytochemical techniqUE for demonstration of DNA in cells infected with mycoplasmas and viruses
應(yīng)用方向:診斷細(xì)胞培養(yǎng)的支原體感染


3.Stage-specific differences in cell cycle control in Trypanosoma brucei revealed by RNA interference of a mitotic cyclin

應(yīng)用方向:流式細(xì)胞術(shù)中細(xì)胞核含量測(cè)定及染色體分選

4.Loss of HSulf-1 up-regulates heparin-binding growth factor signaling in cancer
應(yīng)用方向:評(píng)估細(xì)胞凋亡

5.Direct measurement of femtogram amounts of DNA in cells and chloroplasts by quantitative microspectrofluorometry.
應(yīng)用方向:免疫熒光法和原位雜交法檢測(cè)細(xì)胞核和細(xì)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(yīng)用方向:免疫熒光法和原位雜交法檢測(cè)細(xì)胞核和細(xì)胞器DNA

7.A simple cytochemical techniqUE for demonstration of DNA in cells infected with mycoplasmas and viruses
應(yīng)用方向:取代溴化乙錠對(duì)瓊脂糖凝膠中的DNA進(jìn)行染色

8.Comparison of ethidium bromide and 4', 6'-diamidino-2-phenylindole as quantitative fluorescent stains for DNA in agarose gels
應(yīng)用方向:取代溴化乙錠對(duì)瓊脂糖凝膠中的DNA進(jìn)行染色

9.Localization of polyphosphates in Saccharomyces fragilis, as revealed by 4′, 6-diamidino-2-phenylindole fluorescence
應(yīng)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(jié)合

10.InflUEnce of S-adenosylmethionine on 4′, 6-diamidino-2-phenylindole dihydrochloride (DAPI)-induced fluorescence of polyphosphate in the yeast vacuole
應(yīng)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(jié)合

11.Fluorescent Complexes of DNA With DAPI 4',6-diamidine-2-phenyl indole.2HCl or DCI 4',6-dicarboxyamide-2-phenyl Indole
應(yīng)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(jié)合

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