人微管蛋白抗体的制备及其用于死亡时间推断的价值
Antibody preparation of human tubulin and its value in evaluating postmortem interval

目的：制备人微管蛋白抗体，并用其观察大鼠死亡后肾脏组织中微管蛋白的降解规律与死亡时间的关系。方法：通过分子生物学方法制备小鼠抗人微管蛋白多克隆抗体。取48只大鼠，分为断颈致死组和溺死组，每组24只，采用Western blot观察死亡后24、48、72和96 h大鼠肾脏组织中微管蛋白的降解规律。结果：制备的小鼠抗人微管蛋白多克隆抗体效价超过18 000；2组大鼠死亡后，肾脏组织中微管蛋白均呈逐步降解的趋势；断颈致死组大鼠死后96 h肾脏组织检测不到微管蛋白；溺死组大鼠死后72 h肾脏组织检测不到微管蛋白。结论：大鼠死亡后，肾脏组织中微管蛋白呈逐步降解的趋势；溺死和断颈大鼠死亡后肾脏组织中微管蛋白的降解速度存在差异。
Aim: To prepare human tubulin antibody and explore the relationship between degradation of tubulin in rat kidney tissue and postmortem interval（PMI） after death. Methods: Molecular biology methods were used to purity the mouse anti？？human tubulin polyclonal antibody. A total of 48 rats were allocated into broken neck death group(n=24) and drowning group(n=24). Western blot was used to investigate the degradation of tubulin at 24,48,72,and 96 h after death.Results: The antibody titer of obtained antibodies reached to 18 000. Tubulin could not be detected in rat kidney tissue at 96 h after death in broken neck death group，while could not be detected at 72 h after death in drowning group.Conclusion: The content of tubulin is decreasing with time,which degradates at different speed in rats with different cause of death