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以~3H-苄普地尔作为放射性配体,从牛脑中提取钙调素作为结合受体,采用平衡柱结合法.研究了~3H-苄普地尔与牛脑钙调素的结合特性,建立了~3H-苄普地尔放射受体结合分析法。~3H-苄普地尔仅在Ca~(2+)存在下才能与钙调素产生特异结合,结合具有饱和性和可逆性。Scatchard分析得K_D=2.52±0.44μmol/l,B_(max)=13.95±2.33nmol/mg蛋白,Hill分析得n_H=0.96±0.03。该方法的批内和批间变异系数均为10％,重现性较好。还介绍了使用本法于新设计合成的化合物的竞争性抑制试验初步结果。
Using 3H-bepridil as a radioligand, calmodulin was extracted from bovine brain as a binding partner and the binding properties of ~ 3H-bepridil and bovine calmodulin were studied by a balanced column method , A ~ 3H-bepridil receptor binding assay was established. ~ 3H-bepridil can produce specific binding to calmodulin only in the presence of Ca ~ (2+), with saturation and reversibility. Scatchard analysis gave K_D = 2.52 ± 0.44 μmol / l, B max = 13.95 ± 2.33 nmol / mg protein, and Hill assay gave n_H = 0.96 ± 0.03. The intra-and inter-variation coefficients of this method are both 10%, with good reproducibility. The preliminary results of the competitive inhibition test of the newly designed and synthesized compounds using this method are also introduced.