当含有蛋白质的SDS-PAGE胶浸泡在三氯乙酸或氯仿中时,经三氯化合物的作用,使得蛋白质中的色氨酸经紫外光照射会发出可见光,因而可在SDS-PAGE凝胶上实现对蛋白质条带的定位与定性。免染技术即是基于此原理而发明的。基于免染技术,我们可以检测到待测样品中的总蛋白条带的光密度,进而利用目的蛋白条带光密度与总蛋白条带光密度的比值对目的蛋白进行定量。本文就蛋白质印迹法技术中的免染技术和总蛋白定量方法的原理和优势进行简要综述。
Based on the principle that after UV stimulation,the tryptophan will emit visible light when a SDSPAGEgel containing proteins be soaked in the trichloroacetic acid or chloroform,the stain-free technology was invented.By means of stain-free technology,we can examine the optical density(OD)value of total protein in samples and then,we can quantify the target protein through utilizing the OD value ratios between target protein andtotal proteins.Here,we summarized the principle and advantages of the stain-free technology and the total proteinanalysis method.