产品描述：

保存温度：2-8度

主要优点：

• 无需放射性物质。
• 应用 – 流式细胞仪或荧光显微镜。
• 检测细胞水平的细胞溶解活性。
• 适用于多种类型的哺乳动物细胞系。

技术

测量CMC / ADCCzui常用的方法是放射性铬-51（51Cr）释放试验（2）。该测定有几个缺点：昂贵的，难以加载某些细胞类型，由于严格的环境规定而处置昂贵，并且具有51Cr自发释放的高背景。使用流式细胞仪，现在可以消除对放射性物质的需要，并增加在单个细胞基底上定量细胞溶解活性的能力。各组已经证明通过流式细胞术测量CMC / ADCC活性与传统的51Cr释放测定（3,4,5,6）具有强烈（95％）的相关性。

测定原理

细胞跟踪染料CFSE（7,8,9）用于标记靶细胞群体。在测定完成实验方案后，加入7AAD（活/死）（10,11）以测量细胞死亡。7AAD仅进入膜受损细胞并与DNA结合。

流式细胞术用于门靶细胞并测量7AAD阴性对7AAD后细胞。％细胞毒性通过以下等式计算（参见下面的实验例）：

7AAD正（右上象限）= R1 / 7AAD Postitve（右上象限）= R1 + 7AAD负（右下象限）= R2 ×100 
ACT1

为了测试猪gd淋巴细胞的自然杀伤能力，将K562细胞染色并调整至含有10％FBS的1×10 4个细胞/100μlPRMI的终浓度。以25:1,50:1和100:1的E / T比加入gd淋巴细胞，调节至400μlRPMI的总体积，然后在37℃，无菌封盖的面管中孵育4小时。孵育后，将活/死染色剂直接加入每个管中，在冰上孵育15分钟并通过流式细胞术分析。

引用文献

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