钙离子荧光探针Fluo-8H, AM
货号21091存储条件F/L
规格10x50 ug价格2340
Ex (nm)494Em (nm)517
分子量1074.98溶剂DMSO
产品详细介绍


简要概述

        钙离子荧光探针Fluo-8H, AM是美国AAT Bioquest生产的用于标记钙离子的荧光探针,钙的测量对于许多生物学研究至关重要。荧光探针显示结合Ca2+后的光谱响应,使研究人员能够通过荧光显微镜,流式细胞仪,荧光光谱和荧光酶标仪研究细胞内游离Ca2+浓度的变化。Fluo-3和Fluo-4最常用于可见光可兴奋的钙指示剂中。然而,Fluo-3 AM和Fluo-4 AM在酯酶水解后仅在活细胞中发生中度荧光,并且需要苛刻的细胞负载条件以最大化其细胞钙响应。Fluo-8®染料的开发是为了改善细胞负载和钙响应,同时保持方便的Fluo-3和Fluo-4光谱波长,最大激发波长为~490 nm,最大发射波长为~520 nm。Fluo-8®AM仅需要室温,而Fluo-3 AM和Fluo-4 AM需要37°C的电池负载。此外,Fluo-8®比Fluo-4 AM亮2倍,比Fluo-3 AM亮4倍。AAT Bioquest提供一系列具有不同钙结合亲和力的优异Fluo-8®试剂(Fluo-8®:Kd = 389 nM; Fluo-8H:Kd = 232 nM; Fluo-8L:Kd =1.86μM; Fluo-8FF :Kd =10μM)。我们还提供多种包装尺寸,以满足您的特殊需求,例如1毫克; 10x50μg; 20x50μg; HTS包装,无需额外包装费用。百萤生物是AAT Bioquest 的中国代理商,为您提供最优质的钙离子荧光探针。

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产品说明书

操作步骤

1.准备HHBS缓冲液,10%Pluronic®F-127溶液和25 mM Probenecid溶液。
 

2.在高质量无水DMSO中制备2 mM至5 mM Fluo-8H,AM原液。
2.1使用的Fluo-8H,AM的量:1毫克
2.2所需浓度:2 mM
2.3在合适的容器中,将1mg Fluo-8H TM,AM与465.12μL无水DMSO混合。
 

3.使用10μMFluo-8H ,AM 4在HHBS中制备2X工作溶液,0.08%Pluronic®F-127和2 mM丙磺舒。
3.1最终孔内浓度为Fluo-8H ,AM:5μM
3.2Pluronic®F-127的最终井内浓度:0.04%
3.3最终孔内浓度的丙磺舒:1mM
3.4在合适的容器中混合16μLFluo-8H TM,AM,25.6μL10%Pluronic F-127和256μL25mM丙磺舒。然后,添加HHBS或您选择的缓冲液,直到体积为3.2 mL。
注意:对于大多数细胞系,我们建议使用Fluo-8H™的最终浓度,AM为4至5μM。
注意:推荐的Pluronic F-127井浓度最终为0.02%至0.04%。
注意:推荐的最终浓度为1至2.5 mM的Probenecid。
 

4.将100μL染料工作溶液加入已经含有100μL培养基的所需孔中。
4.1该步骤将染料工作溶液从2X稀释至1X,并将每种组分的最终浓度调节至以下:5μMFluo-8H ,AM,0.04%Pluronic F-127,1mM丙磺舒。
 

5.孵育染料
5.1将染料加载板在细胞培养箱中孵育20-120分钟。
5.2将染料加载板在室温下孵育30分钟。
 

6.用1.0 mM Probenecid准备HHBS缓冲液(或您选择的缓冲液)
6.1在合适的容器中加入160μL的25mM丙磺舒。接下来,添加HHBS或您选择的缓冲液,直到体积为4 mL。
 

7.用HHBS缓冲液或您选择的缓冲液替换染料工作溶液,使用1.0 mM Probenecid。
7.1首先,从所需孔中除去200μL染料工作溶液和培养基。
7.2在相同的孔中加入200μL含有1.0mM丙磺舒的HHBS(或您选择的缓冲液)。
 

8.运行实验
8.1为您的样品添加所需的处理。
8.2以Ex / Em = 494/517 nm运行实验。

 

参考文献

Passive and parallel microfluidic formation of droplet interface bilayers (DIBs) for measurement of leakage of small molecules through artificial phospholipid membranes
Authors: Magdalena A Czekalska, Tomasz S Kaminski, Karol Makuch, Piotr Garstecki
Journal: Sensors and Actuators B: Chemical (2019)

Development of micro mechanical device having two-dimensional array of micro chambers for cell stretching
Authors: K Minami, T Hayashi, K Sato, T Nakahara
Journal: Biomedical microdevices (2018): 10

Spatiotemporal magnetic fields enhance cytosolic Ca 2+ levels and induce actin polymerization via activation of voltage-gated sodium channels in skeletal muscle cells
Authors: Mónica Rubio Ayala, Tatiana Syrovets, Susanne Hafner, Vitalii Zablotskii, Alexandr Dejneka, Thomas Simmet
Journal: Biomaterials (2018)

2-OMe-lysophosphatidylcholine analogues are GPR119 ligands and activate insulin secretion from βTC-3 pancreatic cells: Evaluation of structure-dependent biological activity
Authors: Anna Drzazga, Agata Sowińska, Agnieszka Krzemińska, Andrzej Okruszek, Piotr Paneth, Maria Koziolkiewicz, Edyta Gendaszewska-Darmach
Journal: Biochimica et Biophysica Acta (BBA)-Molecular and Cell Biology of Lipids (2017)

A new electro-optical approach for conductance measurement: an assay for the study of drugs acting on ligand-gated ion channels
Authors: A Menegon, S Pitassi, N Mazzocchi, L Redaelli, R Rizzetto, JF Rolland, C Poli, M Imberti, A Lanati, F Grohovaz
Journal: Scientific Reports (2017)

Altered spontaneous calcium signaling of in situ chondrocytes in human osteoarthritic cartilage
Authors: Xiaoyuan Gong, Wenbin Xie, Bin Wang, Lingchuan Gu, Fuyou Wang, Xiang Ren, Cheng Chen, Liu Yang
Journal: Scientific reports (2017): 17093

Analysis of Ca2+ response of osteocyte network by three-dimensional time-lapse imaging in living bone
Authors: Tomoyo Tanaka, Mitsuhiro Hoshijima, Junko Sunaga, Takashi Nishida, Mana Hashimoto, Naoya Odagaki, Ryuta Osumi, Taiji Aadachi, Hiroshi Kamioka
Journal: Journal of Bone and Mineral Metabolism (2017): 1--10

Aryl-and alkyl-phosphorus-containing flame retardants induced mitochondrial impairment and cell death in Chinese hamster ovary (CHO-k1) cells
Authors: Chao Huang, Na Li, Shengwu Yuan, Xiaoya Ji, Mei Ma, Kaifeng Rao, Zijian Wang
Journal: Environmental Pollution (2017): 775--786

Bystander effects elicited by single-cell photo-oxidative blue-light stimulation in retinal pigment epithelium cell networks
Authors: Masaaki Ishii, Bärbel Rohrer
Journal: Cell Death Discovery (2017): 16071

Ca 2+ signals initiate at immobile IP 3 receptors adjacent to ER-plasma membrane junctions
Authors: Nagendra Babu Thillaiappan, Alap P Chavda, Stephen C Tovey, David L Prole, Colin W Taylor
Journal: Nature Communications (2017): 1505

 

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说明书
钙离子荧光探针Fluo-8H, AM.pdf