高原鼢鼠睾丸雌激素受体免疫组化染色最适浓度抗体的筛选

Effects of different concentrations of antibody on immunohistochemical staining of estrogen receptor in testicle of plateau zokor

摘要:

【目的】筛选高原鼢鼠（Eospalax baileyi）睾丸雌激素受体（Estrogen receptor，ER）免疫组化染色的适宜抗体浓度。【方法】研究选取 10 只成年雄性高原鼢鼠的睾丸，采用免疫组织化学 SP 法 （IHC-SP 法）检测雌激素受体 α（ERα）和雌激素受体 β（ERβ）在 1∶150、1∶200 和 1∶250 稀释浓度下的染色效果。【结果】 ERα 抗体稀释浓度 1∶250 的免疫组化平均光密度值显著高于 1∶150 和 1∶200 的抗体稀释浓度免疫组化平均光密度值（P＜0. 05），而 ERβ 抗体稀释浓度 1∶200 的平均光密度值最高（P＜ 0. 01），最适浓度结果均表现出阳性细胞染色强度高、背景清晰。【结论】高原鼢鼠睾丸中 ERα 和 ERβ 抗体稀释浓度分别为 1∶250 与 1∶200 的免疫组化染色效果最佳。

Abstract:

【Objective】 This research aimed to determine the appropriate antibody concentration for immunohisto- chemical staining of estrogen receptor （ER） in testes of plateau zokor （Eospalax baileyi）.【Method】 We selected the testes of 10 adult male plateau zokors were selected to detect the staining effect of estrogen receptor α（ERα） and es- trogen receptor β（ERβ） at three different dilution concentrations：1∶150，1∶200，and 1∶250 using the immunohisto- chemical SP method （IHC SP method）.【Result】 The mean immunohistochemical optical density of ERα antibody diluted at 1∶250 was significantly higher than that of antibody diluted at 1∶150 and 1∶200（P＜0. 05）. In contrast，the immunohistochemical mean optical density of ERβ antibody diluted at 1∶200 was the highest （P＜0. 01）. At the opti- mal dilution concentration， positive cells exhibited intense staining with a clear background.【Conclusion】 For the tes- tes of plateau zokors， the ideal antibody concentrations for ERα and ERβ were found to be 1∶250 and 1∶200，respec- tively，as these concentrations yielded the best immunohistochemical staining effects.