检测原理 利用 LIVE/DEAD® 细胞成像试剂盒,可通过在活细胞内保存完好的几乎不发荧光的细胞透过性钙黄绿素 AM 和高强度荧光钙黄绿素的酶催化转化,确定细胞内普遍存在的酯酶活性,从而区分活细胞。LIVE/DEAD® 细胞成像试剂盒中的红色组分是细胞不透过性的,因此只能进入具有受损细胞膜的细胞。在即将死亡的和死亡细胞中与 DNA 结合后,会产生明亮的红色荧光。该检测技术本身的背景荧光水平较低,因为染料在与细胞相互作用前几乎不发荧光。基于其中荧光基团的亮度、光谱特性(FITC 和 Texas Red 滤光片)和易用性而选择 LIVE/DEAD® 细胞成像试剂盒。由于其针对工作流程便利性的包装,可轻松快速地测定细胞群中的活细胞和死亡细胞组分。
I need to use a dead cell control for my viability assay. Do you have a protocol for killing cells for this?
Heat killing is commonly used. Place your cells in a tube in buffer and heat at 60oC for 20 minutes. You can also kill your cells by fixing them with ice cold 70% ethanol for 15 minutes. The ethanol-killed cells can then be stored at -20oC until needed, at which point you wash out the ethanol and replace with buffer.
Can I use the LIVE/DEAD Cell Imaging Kit (488/570) (Cat. No. R37601) with fixed cells?
The LIVE/DEAD Cell Imaging Kit (488/570) (Cat. No. R37601) is used for identifying live and dead cells using fluorescence-based staining. It cannot be used with fixed cells.
What is the composition of the Live Green (Component A) of the LIVE/DEAD Cell Imaging Kit (488/570) (Cat. No. R37601)?
The formulation of the solution as well as the amount and concentration of Calcein, AM provided in the LIVE/DEAD Cell Imaging Kit (488/570) (Cat. No. R37601) is proprietary information.