Basile A S, Skolnick P
J Pharmacol Methods. 1987 Apr;17(2):149-56. doi: 10.1016/0160-5402(87)90025-8.
The equilibrium binding constants of [3H]Ro 5-4864 (a "peripheral" benzodiazepine receptor ligand) to renal membranes preserved by various freezing techniques were investigated. The Bmax for [3H]Ro 5-4864 binding to membranes from kidneys preserved as unwashed homogenates stored at -80, -20, or 5 degrees C, whole kidneys stores at -20 or 5 degrees C or as washed homogenate stored at -20 degrees C was significantly decreased (approximately 35%). Only when kidneys were frozen intact (using a dry-ice/acetone slurry) and stored at -80 degrees C was the density of [3H]Ro 5-4864 binding unchanged. However, the Bmax of [3H]PK 11195 (a putative "peripheral" benzodiazepine receptor antagonist) binding to renal membranes was unchanged following storage techniques that reduced the density of [3H]Ro 5-4864 binding 38%. No change was observed in the Kd values for [3H]Ro 5-4864 and [3H]PK 11195 binding to renal membranes preserved under any condition tested. These results demonstrate a method for the preservation of [3H]Ro 5-4864 binding to renal membranes, and suggests that [3H]Ro 5-4864 and [3H]PK 11195 bind to unique sites on or near the "peripheral" benzodiazepine receptor. [corrected]
研究了[3H]Ro 5-4864（一种“外周”苯二氮䓬受体配体）与通过各种冷冻技术保存的肾膜的平衡结合常数。[3H]Ro 5-4864与保存在-80℃、-20℃或5℃的未洗涤匀浆、保存在-20℃或5℃的完整肾脏或保存在-20℃的洗涤匀浆中的肾脏膜结合的Bmax显著降低（约35%）。只有当肾脏完整冷冻（使用干冰/丙酮浆液）并保存在-80℃时，[3H]Ro 5-4864结合的密度才保持不变。然而，[3H]PK 11195（一种假定的“外周”苯二氮䓬受体拮抗剂）与肾膜结合的Bmax在降低[3H]Ro 5-4864结合密度38%的保存技术后没有变化。在任何测试条件下保存的肾膜上，[3H]Ro 5-4864和[3H]PK 11195结合的Kd值均未观察到变化。这些结果证明了一种保存[3H]Ro 5-4864与肾膜结合的方法，并表明[3H]Ro 5-4864和[3H]PK 11195与“外周”苯二氮䓬受体上或其附近的独特位点结合。[已校正]
