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細胞膜熒光探針DiD標記液

貨號22033存儲條件 在零下15度以下保存, 避免光照
規格10 mL價格2544
Ex (nm)646Em (nm)663
分子量959.91溶劑DMSO
產品詳細介紹


簡要概述

細胞膜熒光探針DiD標記液是美國AAT Bioquest生產的用于細胞膜檢測的熒光探針,DiA,DiI,DiO,DiD和DiR染料是用于標記細胞膜和其他疏水結構的親脂性熒光染料家族。當摻入膜中或與親脂性生物分子如蛋白質結合時,這些對環境敏感的染料的熒光大大增強,盡管它們在水中是弱熒光的。它們具有高消光系數,極性依賴性熒光和短激發態壽命。一旦應用于細胞,這些染料在細胞質膜內橫向擴散,導致整個細胞以其最佳濃度均勻染色。 DiI(橙色熒光),DiO(綠色熒光),DiD(紅色熒光)和DiR(深紅色熒光)的獨特熒光顏色為活細胞的多色成像和流式細胞分析提供了便利的工具。 DiO和DiI可分別與標準FITC和TRITC過濾器一起使用。其中DiD受633 nm He-Ne激光器的激發,并且具有比DiI更長的激發和發射波長,為標記具有顯著內在熒光的細胞和組織提供了有價值的替代方案。 DiR可能對體內成像或追蹤有用,因為紅外光通過細胞和組織的有效傳播和低水平的紅外線范圍內的自發熒光。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DiD標記液。 

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產品說明書

操作方法

1.準備DiO,DiI,DiD,DiS或DiR膜染色溶液:

1.1制備DMSO或EtOH儲備溶液:儲備溶液應在DMSO或EtOH中以1-5mM制備。

注意:儲備溶液的未使用部分應儲存在-20 ℃。 避免反復凍/融循環。

1.2準備工作溶液:將儲備溶液(步驟1.1)稀釋到合適的緩沖液中,如無血清培養基,HBSS或PBS制備1至5μM的工作溶液。

注意:對于不同的細胞類型和/或實驗條件,應根據經驗確定工作溶液的最終濃度。 建議在至少超過十倍范圍的濃度下進行測試。

 

2.將細胞染成懸浮液:

2.1在染料工作溶液中懸浮細胞密度為1×106 / mL(來自步驟1.2)。

2.2在37°C孵育2-20分鐘。 最佳孵育時間取決于細胞類型。 首先孵育20分鐘,然后根據需要進行優化以獲得均勻的標記。

2.3將標記的懸浮管以1000至1500rpm離心5分鐘。

2.4取出上清液,輕輕地將細胞重新懸浮在預熱(37°C)的生長培養基中。

2.5按步驟2.3和2.4洗滌兩次。

 

3.染色貼壁細胞:

3.1在無菌玻璃蓋玻片上培養貼壁細胞。

3.2從生長培養基中取出蓋玻片,輕輕地排出多余的培養基。 將蓋玻片放在濕度箱中。

3.3將100μL染料工作溶液(來自步驟1.2)吸移到蓋玻片的角落,輕輕攪拌直至所有細胞都被覆蓋。

3.4將蓋玻片在37°C孵育2-20分鐘。 最佳孵育時間取決于細胞類型。 首先孵育20分鐘,然后根據需要進行優化以獲得均勻的標記。

3.5排出染料工作溶液,用生長培養基清洗蓋玻片2-3次。每個洗滌循環用預熱的生長培養基覆蓋細胞,孵育5-10分鐘后排出培養基。

 

4.顯微鏡檢測:

4.1說明書中的表1總結了DiD,DiO,DiI,DiS和DiR濾波器組的選擇。

4.2為了同時檢測多種染料,可提供如下多波段濾波器組:

a)DiI和DiO = Omega XF52,Chroma 51004

b)DiI和DiD = Omega XF92,Chroma 51007

c)DiI,DiO和DiD = Omega XF93,Chroma 61005

 

5.流式細胞儀檢測:

用DiO,DiI,DiD,DiS和DiR標記的細胞可分別使用常規FL1,FL2,FL3和FL4流式細胞儀檢測通道進行分析。

 

參考文獻

Exosomes from hyperglycemia-stimulated vascular endothelial cells contain versican that regulate calcification/senescence in vascular smooth muscle cells
Authors: Shuang Li, Jun-Kun Zhan, Yan-Jiao Wang, Xiao Lin, Jia-Yu Zhong, Yi Wang, Pan Tan, Jie-Yu He, Xing-Jun Cui, Yi-Yin Chen
Journal: Cell & Bioscience (2019): 1

The Influence of Cell Source and Donor Age on the Tenogenic Potential and Chemokine Secretion of Human Mesenchymal Stromal Cells
Authors: Weronika Zarychta-Wisniewska, Anna Burdzińska, Katarzyna Zielniok, Marta Koblowska, Kamila Gala, Piotr Pedzisz, Roksana Iwanicka-Nowicka, Anna Fogtman, Aleksandra Aksamit, Agnieszka Kulesza
Journal: Stem Cells International (2019)

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)



說明書
細胞膜熒光探針DiD標記液.pdf


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