钙黄绿素绿，AM

钙黄绿素绿，AM是美国AAT Bioquest生产的用于检测活细胞的荧光探针，Calcein UltraGreen AM可轻松通过活细胞的细胞膜。在运输到活细胞中时，细胞酯酶切断了AM基团，该分子被捕获在细胞内。与Calcein AM相比，Calcein UltraGreen 更适合用于染色活细胞的荧光探针，因为它具有较低的细胞毒性和较长的细胞保留时间。UltraGreen AM不会显着影响细胞功能，如淋巴细胞的增殖或趋化性。百萤生物是AAT Bioquest的中国代理商，为您提供优质的钙黄绿素绿，AM。 

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选择指南

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 Calcein UltraGreen AM原液。
2.1使用Calcein UltraGreen 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钙黄绿素UltraGreen TM AM与823.09μL无水DMSO混合。
 

3.使用10μMCalceinUltraGreen 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终孔内浓度的Calcein UltraGreen AM：5μM
3.2Pluronic®F-127的终孔内浓度：0.04％
3.3终孔内浓度的丙磺舒：1mM
3.4在合适的容器中混合16μL的Calcein UltraGreen AM，25.6μL的10％Pluronic®F-127和256μL的25 mM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Calcein UltraGreen AM的终浓度为4至5μM。
注意：推荐的Pluronic F-127井浓度终为0.02％至0.04％。
注意：推荐的终浓度为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，并将每种组分的终浓度调节至以下：5μM钙黄绿素UltraGreen 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20-60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 = 496/524 nm运行实验。

参考文献

Functional imaging of neuronal activity of auditory cortex by using Cal-520 in anesthetized and awake mice
Authors: Jingcheng Li, Jianxiong Zhang, Meng Wang, Junxia Pan, Xiaowei Chen, Xiang Liao
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NINJ2--A novel regulator of endothelial inflammation and activation
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Erythropoietin Stimulates Endothelial Progenitor Cells to Induce Endothelialization in an Aneurysm Neck After Coil Embolization by Modulating Vascular Endothelial Growth Factor
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Flexible Endoscopic Spray Application of Respiratory Epithelial Cells as Platform Technology to Apply Cells in Tubular Organs
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Rapid generation of collagen-based microtissues to study cell--matrix interactions
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Journal: Journal of Applied Toxicology (2016)

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