兔抗褪黑激素高免血清的制备、提取和纯化  

PREPARATION EXTRACT AND PURIFICATION OF RABBIT ANTI\|MELATONIN ANTIBODIES

作　　者：汪兴生[1] 尚平[2] 姜勋平[1] 夏银 王进荣[1] 程瑞禾[1] 韦来红[1] 

机构地区：[1]南京农业大学动物科技学院,江苏南京210095 [2]南京军区总医院,江苏南京210000 [3]新英格兰大学物理系

出　　处：《江苏农业研究》2000年第1期29-32,共4页

基　　金：江苏省自然科学资金!资助项目 ( BK970 95 )

摘　　要：运用 Mannich法 ,将褪黑激素 ( MLT)与牛血清白蛋白 ( BSA)偶联制备成完全抗原。经 UV-2 60紫外分光光度仪测定 ,MLT与 BSA的连接比为 1 2∶ 1。 5只新西兰大耳兔用此抗原免疫 6次后 ,ELISA和 RIA抗体效价分别平均 1∶1 40 0 0和 1∶ 1 50 0 0 0。抗血清经 Na2 SO4沉淀和 Sephedex-G-2 0 0层析后收集纯化抗体 ,用 72 1分光光度计测其蛋白浓度。通过Melatonin complete antigen was prepared through Mannich′s formaldehyde condensation reaction with melatonin to BSA molar ratio of 12∶1.5 New Zealand white rabbits were immunized using the antigen. After the 6 booster injections, the average titers of ELISA and RIA were 1∶14 000 and 1∶150 000, respectively. The antiserum were precipitated with Na\-2SO\-4 and passed through Sephedex\|G\|200 columus. The purified antibody was collected and detected by UV\|721 table spectrophotometry. The cross\|reaction with 5\|methoxy\|tryptophan, a derivative of melatonin, showed that the antibody was melatonin\|specific.

关 键 词：褪黑激素 抗原 抗体 BSA 制备 提取 纯化 兔 

分 类 号：Q576.03[生物学—生物化学] S852.5[农业科学—基础兽医学]