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本文报道利用高度敏感的亲和素-生物素过氧化物酶(ABC)试剂,建立一种测定体外合成抗体的ELISA.将抗原致敏动物的脾脏细胞悬液直接加在包盖过抗原的微量滴定板孔内,置37℃的CO_2培养箱内温育6小时后,洗去细胞,各孔加生物素化第二抗体,温育2小时后洗板,继加ABC,反应30分钟后洗板,加底物显色.若在细胞悬液中加入蛋白合成抑制剂嘌呤霉素,可使光密度读数下降,从而证明参加反应的抗体确系体外合成.此法测定重复样品的
This article reports the use of a highly sensitive avidin-biotin peroxidase (ABC) reagent to establish an ELISA for the in vitro synthesis of antibodies. The antigen-sensitized animal spleen cell suspension was added directly to the antigen Titration plate wells, set 37 ℃ CO_2 incubator incubated for 6 hours, wash away cells, biotinylated secondary antibody with each well, incubated for 2 hours after the wash plate, followed by ABC, the reaction after 30 minutes wash Plate, add substrate color.If adding protein synthesis inhibitor puromycin in the cell suspension, the optical density readings can be reduced, thus proving that the reaction to participate in the reaction of the antibody is indeed in vitro synthesis. This method for the determination of repeated samples