放射配基法测定大鼠海马μ-阿片受体的方法总结及优化  

RADIOLIGAND BINDING ASSAY OPTIMIZATION FOR DETECTING THE μ-OPIOID RECEPTOR IN HIPPOCAMPUS OF RATS

作　　者：文迪[1] 丛斌[1] 马春玲[1] 张雅静[1] 于峰[1] 

机构地区：[1]河北医科大学基础医学院法医学系河北省法医学重点实验室,河北石家庄050017

出　　处：《河北医科大学学报》2009年第4期325-327,共3页Journal of Hebei Medical University

基　　金：国家自然科学基金(30672355);河北省自然科学基金(C2007000826)

摘　　要：目的建立一套安全、经济、可靠的实验条件，为检测大鼠海马“阿片受体奠定方法学基础。方法从膜蛋白提取、结合反应体系、孵育温度、孵育时间等方面，对放射配基结合分析的实验条件进行摸索；并从环保、经济的角度考虑，对实验进行了优化。结果发现当蛋白浓度为1mg／mL、[^2H]DAMGO 0．5～8．0μmol／L5个浓度点、非标记配基DAMGO浓度为5μmol／L、CC孵育过夜的条件下，结果稳定、可靠。结论该方法经济、可靠、重复性好，且易于掌握、污染少。Objective To establish a safe, sparing and credible experimental technique for detecting the μ-opioid receptor in hippocampus of rats. Methods Optimizing radioligand binding assay conditions, such as the process of protein extraction, the condition of the binding reaction, incubate temperature and time, etc. and making it more sparing and less contamination. Results The result was excellent when the protein concentration was 1 mg/mL, [^3H]DAMGO concentrations were among 0.5-8.0 nmol/L and unlabled DAMGO concentration was 5μmol/L, 4℃ overnight. Conclusion The protocol is sparing, reliable and reproclucible. The method is easy to master and less contamination.

关 键 词：放射性同位素 受体 阿片样 μ 大鼠 

分 类 号：R817.1[医药卫生—影像医学与核医学]