Treherne J M, Young J M
Department of Pharmacology, University of Cambridge, UK.
J Pharm Pharmacol. 1988 Oct;40(10):730-3. doi: 10.1111/j.2042-7158.1988.tb07006.x.
The binding of [3H]mepyramine to histamine H1-receptors solublized from guinea-pig cerebellum by 1% digitonin could be assayed by adsorption of the receptor-bound [3H]ligand to glass-fibre filters pretreated with 0.3% polyethylenimine (PEI). Non-specific binding was higher than in parallel experiments in which the bound and free [3H]ligand was separated by gel-filtration on Sephadex G-25 columns, but the parameters characterizing the inhibition curves were otherwise similar. The PEI-treated filter method could also be used to assay [3H]-(+)-N-methyl-4-methyl-diphenhydramine ([3H]QMDP) binding to solubilized H1-receptors, but the level of non-specific binding was higher and was only satisfactorily defined by icotidine or temelastine. A particular utility of the PEI-treated filter assay will be in measurements of the kinetics of H1-receptor-ligand interactions.
通过1%洋地黄皂苷从豚鼠小脑中溶解出的组胺H1受体与[3H]美吡拉敏的结合，可通过将受体结合的[3H]配体吸附到用0.3%聚乙烯亚胺（PEI）预处理的玻璃纤维滤膜上来进行测定。非特异性结合高于通过在Sephadex G - 25柱上进行凝胶过滤分离结合态和游离态[3H]配体的平行实验，但表征抑制曲线的参数在其他方面相似。经PEI处理的滤膜法也可用于测定[3H] -（+）-N - 甲基 - 4 - 甲基 - 二苯海拉明（[3H]QMDP）与溶解的H1受体的结合，但非特异性结合水平更高，且仅通过异可替丁或替美司丁才能令人满意地界定。经PEI处理的滤膜测定法的一个特殊用途将在于测量H1受体 - 配体相互作用的动力学。
