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細胞膜熒光探針DiA

貨號22030存儲條件 在零下15度以下保存, 避免光照
規格25 mg價格984
Ex (nm)492Em (nm)613
分子量787.04溶劑DMSO
產品詳細介紹


簡要概述

DiA是一種親脂性熒光染色劑,用于標記膜和其他疏水結構。它具有環境敏感的熒光,盡管在水中呈弱熒光,但與膜或親油生物分子(如蛋白質)結合時,熒光會大大增強。一旦應用于細胞,這種染料在細胞質膜內橫向擴散,從而在最佳濃度下對整個細胞進行均勻染色。百螢生物是AAT Bioquest 的中國代理商,為您提供最優質的細胞膜熒光探針DiA。

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產品說明書

樣品分析方案

概述

用測試化合物(200μL/樣品)制備細胞

添加膜聯蛋白V綴合物測定溶液

室溫孵育30-60分鐘

用流式細胞儀或熒光顯微鏡分析

 

操作步驟

1.用膜聯蛋白V綴合物制備和孵育細胞:

1.1制備膜聯蛋白V結合測定緩沖液:10mM HEPES,140mM NaCl和2.5mM CaCl 2,pH 7.4。

1.2用測試化合物處理細胞一段所需的時間(用星形孢菌素處理的Jurkat細胞4-6小時)以誘導細胞凋亡。

1.3離心細胞,得到1-5×105個細胞/管。

1.4將細胞重懸于200μL膜聯蛋白V結合測定緩沖液中(來自步驟1.1)。

1.5在細胞中加入2μL膜聯蛋白V結合物。

可選:在細胞內加入死細胞染色劑如碘化丙錠,用于壞死細胞。

1.6在室溫下孵育30至60分鐘,避光。

1.7在用流式細胞儀或熒光顯微鏡分析細胞之前,加入300μL膜聯蛋白V結合測定緩沖液(來自步驟1.1)以增加體積(參見下面的步驟1.8)。

1.8使用流式細胞儀或熒光顯微鏡監測熒光強度(參見下面的步驟2或3)。

 

2.使用流式細胞儀分析:

通過使用具有適當過濾器的流式細胞儀來量化膜聯蛋白V綴合物。

注意:粘附細胞上的膜聯蛋白V結合流式細胞術分析未經常檢測,因為在細胞分離或收獲期間可能發生特定的膜損傷。 然而,Casiola-Rosen等人先前報道了利用膜聯蛋白V對貼壁細胞類型進行流式細胞術的方法。 和van Engelend等人(見參考文獻1和2)。

 

3.使用熒光顯微鏡分析:

3.1移取步驟1.6的細胞懸液,用膜聯蛋白V結合測定緩沖液(來自步驟1.1)沖洗1-2次,然后用膜聯蛋白V結合測定緩沖液(來自步驟1.1)重懸細胞。 將細胞添加到蓋有玻璃蓋玻片的載玻片上。

注意:對于粘附細胞,建議直接在蓋玻片上生長細胞。 與膜聯蛋白V綴合物孵育(步驟1.6)后,用膜聯蛋白V結合測定緩沖液(來自步驟1.1)沖洗1-2次,并將膜聯蛋白V結合測定緩沖液(來自步驟1.1)加回到蓋玻片中。 在玻璃載玻片上翻轉蓋玻片并觀察細胞。 在與膜聯蛋白V綴合物孵育后,細胞也可以在2%甲醛中固定,并在顯微鏡下觀察。

3.2在熒光顯微鏡下用適當的濾光片分析膜聯蛋白V綴合物的凋亡細胞。

 

參考文獻

Cyclic RGD peptide-modified liposomal drug delivery system for targeted oral apatinib administration: enhanced cellular uptake and improved therapeutic effects
Authors: Zhiwang Song, Yun Lin, Chan Feng Xia Zhang, Yonglin Lu, Yong Gao, Chunyan Dong
Journal: International Journal of Nanomedicine (2017): 1941

In vivo imaging system for explants analysis—A new approach for assessment of cell transplantation effects in large animal models
Authors: Weronika Zarychta-Wisniewska, Anna Burdzinska, Radoslaw Zagozdzon, Bartosz Dybowski, Marta Butrym, Zdzislaw Gajewski, Leszek Paczek
Journal: PloS one (2017): e0184588

Influence of Particle Geometry on Gastrointestinal Transit and Absorption following Oral Administration
Authors: Dong Li, Jie Zhuang, Haisheng He, Sifan Jiang, Amrita Banerjee, Yi Lu, Wei Wu, Samir Mitragotri, Li Gan, Jianping Qi
Journal: ACS Applied Materials & Interfaces (2017)

Mesenchymal stem cells increase skin graft survival time and up-regulate PD-L1 expression in splenocytes of mice
Authors: Ali Moravej, Bita Geramizadeh, Negar Azarpira, Amir-Hasan Zarnani, Ramin Yaghobi, Mehdi Kalani, Maryam Khosravi, Amin Kouhpayeh, Mohammad-Hossein Karimi
Journal: Immunology Letters (2017)

Novel approach for the detection of intraperitoneal micrometastasis using an ovarian cancer mouse model
Authors: Ayesha B Alvero, Dongin Kim, Eydis Lima, Natalia J Sumi, Jung Seok Lee, Carlos Cardenas, Mary Pitruzzello, Dan-Arin Silasi, Natalia Buza, Tarek Fahmy
Journal: Scientific Reports (2017)

On-Demand Drug Releasing from Dual Targeting Small Nanoparticles Triggered by High Intensity Focused Ultrasound Enhanced Glioblastoma Targeting Therapy
Authors: Zimiao Luo, Kai Jin, Qiang Pang, shun shen, Zhiqiang Yan, Ting Jiang, Xiaoyan Zhu, Lei Yu, Zhiqing Pang, Xinguo Jiang
Journal: ACS Applied Materials & Interfaces (2017)

Overexpression of c-Met in bone marrow mesenchymal stem cells improves their effectiveness in homing and repair of acute liver failure
Authors: Kun Wang, Yuwen Li, Tiantian Zhu, Yongting Zhang, Wenting Li, Wenyu Lin, Jun Li, Chuanlong Zhu
Journal: Stem Cell Research & Therapy (2017): 162

The accelerated blood clearance phenomenon of PEGylated nanoemulsion upon cross administration with nanoemulsions modified with polyglycerin
Authors: Yuqing Su, Lirong Wang, Kaifan Liang, Mengyang Liu, Xinrong Liu, Yanzhi Song, Yihui Deng
Journal: Asian Journal of Pharmaceutical Sciences (2017)

A dual brain-targeting curcumin-loaded polymersomes ameliorated cognitive dysfunction in intrahippocampal amyloid-β1--42-injected mice
Authors: Tingting Jia, Zhiguo Sun, Ying Lu, Jie Gao, Hao Zou, Fangyuan Xie, Guoqing Zhang, Hao Xu, Duxin Sun, Yuan Yu
Journal: International Journal of Nanomedicine (2016): 3765

Annonaceous acetogenins nanosuspensions stabilized by PCL-PEG block polymer: significantly improved antitumor efficacy
Authors: Jingyi Hong, Yanhong Li, Yijing Li, Yao Xiao, Haixue Kuang, Xiangtao Wang
Journal: International Journal of Nanomedicine (2016): 3239



說明書
細胞膜熒光探針DiA.pdf


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