Jamil Hina, Saleemuddin Mohammed
Department of Biochemistry, Faculty of Life Sciences, Aligarh Muslim University, Aligarh 202 002, India.
Indian J Biochem Biophys. 2002 Aug;39(4):274-8.
A simple strategy to remarkably increase the sensitivity of detection of antigens applied as dot or western blot on nitrocellulose membrane using human serum albumin as model antigen has been described. This involves subjecting the antigen bearing nitrocellulose strips to multiple incubation cycles with primary antibody and enzyme conjugated secondary antibody prior to staining for enzyme activity. The sensitivity of detection could be increased up to a thousand fold after three incubation cycles. Aggregation of human serum albumin could be detected by the multiple incubation procedure at very low protein concentration after electrophoresis and transfer onto nitrocellulose.
已描述了一种简单策略，该策略以人血清白蛋白作为模型抗原，可显著提高在硝酸纤维素膜上进行斑点或蛋白质免疫印迹时抗原检测的灵敏度。这包括在对酶活性进行染色之前，使带有抗原的硝酸纤维素条带与一抗和酶偶联二抗进行多次孵育循环。经过三个孵育循环后，检测灵敏度可提高至原来的一千倍。在电泳并转移至硝酸纤维素膜上后，通过多次孵育程序可在极低的蛋白质浓度下检测到人血清白蛋白的聚集。
