McLachlan R
Pathology Department, Peter MacCallum Cancer Institute, Melbourne, Australia.
Clin Chem. 1989 Mar;35(3):478-81.
I describe a simple, economical technique for identifying low concentrations of monoclonal immunoglobulins in the presence of excessive amounts of immunoglobulins of other classes. The technique involves binding of specific antibody to nitrocellulose, separating proteins by isoelectric focusing or zone electrophoresis in agarose gels, using capillary transfer to bind proteins to the nitrocellulose via their antibody affinity, and then detecting transferred proteins with enzyme-labeled antibody. A monoclonal immunoglobulin can be completely characterized in 2 h. No expensive equipment is required. Affinity blotting is about 10-fold as sensitive as native blotting, 100-fold as sensitive as silver staining.
我描述了一种简单、经济的技术，用于在存在过量其他类免疫球蛋白的情况下鉴定低浓度的单克隆免疫球蛋白。该技术包括将特异性抗体结合到硝酸纤维素上，通过等电聚焦或琼脂糖凝胶区带电泳分离蛋白质，利用毛细管转移通过抗体亲和力将蛋白质结合到硝酸纤维素上，然后用酶标记抗体检测转移的蛋白质。单克隆免疫球蛋白可在2小时内完全鉴定出来。无需昂贵设备。亲和印迹的灵敏度约为天然印迹的10倍，银染的100倍。
