DiI标记脂蛋白改良法的建立

The Modification on Lipoprotein Lebeling Method with DiI

为了改进脂蛋白的荧光标记方法 ,本实验采用经超速离心后的离心管底层血清取代商品化的去脂血清作为脂蛋白标记时的介质 ,与荧光染料DiI混合孵育。经超速离心分离后获得DiI标记的极低密度脂蛋白、低密度脂蛋白及β 极低密度脂蛋白。本方法在标记过程中不用去脂血清 ,降低了标记成本并缩短了超速离心时间。配体与受体结合实验结果发现 ,用本方法标记的脂蛋白能以可饱和方式结合体外培养的小鼠腹腔巨噬细胞和中国仓鼠卵母细胞。表明本方法标记的脂蛋白保持了正常脂蛋白的配体功能 。

Aim To get a rapid, simple label method for lipop rotein , an improved method was established with the fluorescent probe DiI. Method The serum at the bottom of the post ultracentrifuged tu be, instead of lipoprotein deficient serum (LPDS), was used as the medium to app lying in lipoprotein labeling. Results Binding assay showed that the labeled lipoprotein can bind with macrophage and chinese hamster ovocyte (CHO) cells with typical saturation able model. The results p roved that present method labeling lipoprotein have the normal lipoprotein ligan d binding properties. Conclusion This method is sim ple, rapid and reliable. It could be applied widely in lipoprotein receptor s tudy.