Cell Meter™细胞粘附测定试剂盒 货号23010-AAT Bioquest荧光染料

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Cell Meter™细胞粘附测定试剂盒

Cell Meter™细胞粘附测定试剂盒

Cell Meter™细胞粘附测定试剂盒    货号23010 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 4272
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。

点击查看光谱

 

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

钙黄绿素超绿AM储备溶液

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

钙黄绿素超绿AM工作溶液

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。

  1. 在涂有所需试剂的板上添加100 µL体积的细胞。
  2. 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。
  3. 除去生长培养基和未贴壁的细胞。
  4. 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。
  5. 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。
  6. 在孔中加入100 µL粘附测定缓冲液。
  7. 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

Cell Meter™细胞粘附测定试剂盒    货号23010

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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说明书
Cell Meter™细胞粘附测定试剂盒.pdf