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Cell Meter™细胞粘附测定试剂盒
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货号 | 23010 | 存储条件 | 在零下15度以下保存, 避免光照 |
| 规格 | 100 Tests | 价格 | 4368 | |
| Ex (nm) | 494 | Em (nm) | 514 | |
| 分子量 | 溶剂 | |||
| 产品详细介绍 | ||||
简要概述
Cell Meter 细胞粘附测定试剂盒是一种快速灵敏的测定方法，用于测量多种细胞类型的细胞-细胞或细胞表面粘附力。在该测定中，将细胞用钙黄绿素UltraGreen AM标记并使其贴壁。去除非贴壁细胞后，使用钙黄绿素UltraGreen的荧光来计算贴壁细胞的数量。使用我们的荧光染料Calcein UltraGreen AM，可以提供一种快速而灵敏的方法来测量多种细胞类型的细胞粘附性。钙黄绿素UltraGreen AM是无荧光的，但一旦加载到细胞中，就会被内源性酯酶裂解，产生高荧光的钙黄绿素UltraGreen，这是一种荧光强，不依赖pH值的细胞质细胞标记，对细胞粘附过程的干扰最小。Cell Meter™细胞粘附测定法设计用于荧光酶标仪。
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适用仪器
| 荧光显微镜 | |
| 激发： | FITC滤波片组 |
| 发射： | FITC滤波片组 |
| 推荐孔板： | 黑色透明 |
| 荧光酶标仪 | |
| 激发： | 490nm |
| 发射： | 525nm |
| cutoff: | 515nm |
| 推荐孔板： | 黑色透明 |
产品说明书
样品分析方案
概述
添加细胞
在37°C下孵育细胞使其贴壁
去掉未贴壁细胞
添加Calcein Ultragreen AM工作溶液
在37°C下孵育细胞20-30分钟
去除上清液并用HHBS或DPBS洗涤细胞
使用荧光酶标仪（Ex / Em = 490/525 nm）检测荧光强度
储备溶液配制
向Calcein Ultragreen AM（组分A）中加入50 µL DMSO（组分C）并充分混合。
注意：将未使用的Calcein Ultragreen AM储备溶液分装在-20°C下，一次性使用，以免冻融循环。
工作溶液配制
将50 µL钙黄绿素Ultragreen AM储备溶液添加到10 mL粘附测定缓冲液中，并充分混合。
注意：不应储存Calcein Ultragreen AM工作溶液，应立即使用。
注意：10 mL Calcein Ultragreen AM工作溶液足以进行100次测试。
操作步骤
以下方案可用作指导，应根据需要进行优化。
- 在涂有所需试剂的板上添加100 µL体积的细胞。
- 在37°C下孵育平板2至3小时。
注意：对于每种细胞系，最佳孵育时间应通过实验进行测试。 - 除去生长培养基和未贴壁的细胞。
- 加入100 µL钙黄绿素Ultragreen AM工作溶液，并在37°C下孵育平板20-30分钟。
注意：对于每种细胞系，最佳孵育时间应通过实验进行测试。 - 除去染料工作溶液，并用1X Hank盐溶液和20 mM Hepes缓冲液或DPBS洗涤细胞一次。
- 在孔中加入100 µL粘附测定缓冲液。
- 使用荧光酶标仪在Ex / Em = 490/525 nm（截止= 515 nm）处检测荧光强度。
图示
图1.在荧光酶标仪中使用Cell Meter™细胞粘附测定试剂盒测量的细胞粘附。将不同程度的Jurkat细胞在加有不同试剂的孔中孵育，然后在37°C下用Calcein Ultragreen AM染色30分钟。在Ex / Em = 490/525 nm处检测信号。
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