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为了筛选植物组织培养的柴胡皂甙a(SSA)的高产量株、研究SSA的体内动态以及用于评价汉方制剂的质量,开发了SSA的ELISA法。 以高碘酸法将SSA与BSA结合(1分子BSA结合SSA7.8分子),用这种SSA-BSA结合物免疫家兔,得抗血清。以所得的抗血清进行高敏感间接ELISA法的开发。固相吸附SSA-OVA结合物,用含有0.1％酪蛋白的PBS,在停止振荡后加120000倍稀释的抗血清与受试药物,37℃培养2h。洗涤固相后,加
In order to screen high-yield strains of saikosaponin a (SSA) in plant tissue culture, to study the in vivo dynamics of SSA, and to evaluate the quality of Kampo preparations, an SSA ELISA method was developed. SSA was combined with BSA by periodic acid method (1 molecule of BSA bound to SSA 7.8 molecule) and the rabbit was immunized with this SSA-BSA conjugate to obtain antiserum. Development of a highly sensitive indirect ELISA method using the obtained antiserum. The SSA-OVA conjugate was immobilized on a solid phase and the antiserum diluted with 120,000 times was added to the test drug with PBS containing 0.1% casein and stopped after the shaking, followed by incubation at 37C for 2 hours. After washing the solid phase, add