细胞培养检测 Pluronic F

操作说明

注意：以下是我们推荐的方案。 该方案仅提供指南，实际应根据您的具体需求进行修改。

1.将10％Pluronic F-127储备溶液稀释到细胞上样缓冲液（如Hanks和20 mM Hepes缓冲液（HHBS）中，稀释度为1：1000至1：500，得到0.02至0.04％的工作溶液。

2.然后将AM酯的DMSO储备溶液稀释到0.02至0.04％的工作溶液（来自步骤1）中，以获得1μM至10μM的最终AM酯浓度。

注意：Pluronic F-127的最终浓度通常保持在0.08％或以下。

3.将细胞在所需温度下孵育10分钟至1小时。

注意：通常，希望使用所需的最少量的AM酯来获得足够的荧光信号噪声水平。

4.标记后，在开始实验之前用HHBS或新鲜培养基洗涤细胞。

参考文献

3D printed lattices as an activation and expansion platform for T cell therapyAuthors: Bahman Delalat, Frances Harding, Batjargal Gundsambuu, Elena M De-Juan-Pardo, Felix M Wunner, Marie-Luise Wille, Marek Jasieniak, Kristen AL Malatesta, Hans J Griesser, Antonio SimulaJournal: Biomaterials (2017)

GD2-specific CAR T Cells Undergo Potent Activation and Deletion Following Antigen Encounter but can be Protected From Activation-induced Cell Death by PD-1 BlockadeAuthors: Tessa Gargett, Wenbo Yu, Gianpietro Dotti, Eric S Yvon, Susan N Christo, John D Hayball, Ian D Lewis, Malcolm K Brenner, Michael P BrownJournal: Molecular Therapy (2016)

Inflammasome components ASC and AIM2 modulate the acute phase of biomaterial implant-induced foreign body responsesAuthors: Susan N Christo, Kerrilyn R Diener, Jim Manavis, Michele A Grimbaldeston, Akash Bachhuka, Krasimir Vasilev, John D HayballJournal: Scientific reports (2016)

Hybrid core/shell microparticles and their use for understanding biological processesAuthors: Akash Bachhuka, Susan N Christo, Alex Cavallaro, Kerrilyn R Diener, Agnieszka Mierczynska, Louise E Smith, Romeo Marian, Jim Manavis, John D Hayball, Krasimir VasilevJournal: Journal of colloid and interface science (2015): 9--17

Scrutinizing calcium flux oscillations in T lymphocytes to deduce the strength of stimulusAuthors: Susan N Christo, Kerrilyn R Diener, Robert E Nordon, Michael P Brown, Hans J Griesser, Krasimir Vasilev, Farid C Christo, John D HayballJournal: Scientific reports (2015): 7760

Individual and population quantitative analyses of calcium flux in T-cells activated on functionalized material surfacesAuthors: Susan N Christo, Ghafar T Sarvestani, Stefani S Griesser, Bryan R Coad, Hans J Griesser, Krasimir Vasilev, Michael P Brown, Kerrilyn R Diener, John D HayballJournal: Australian Journal of Chemistry (2012): 45--49

Solid-state capture and real-time analysis of individual T cell activation via self-assembly of binding multimeric proteins on functionalized materials surfacesAuthors: Kerrilyn R Diener, Susan N Christo, Stefani S Griesser, Ghafar T Sarvestani, Krasimir Vasilev, Hans J Griesser, John D HayballJournal: Acta biomaterialia (2012): 99--107

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