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本文用~3H-QNB作特异结合配基,用简单灵敏的放射配体法测定鼠心肌M受体。经28000×g低温离心制备膜制剂,在2ml Na-K磷酸缓冲液(pH7.4)中含0.8mg膜蛋白,不同浓度~3H-QNB 37℃温育半小时,用国产79型滤膜分离结合与游离部份,用0.01μM QNB作测非特异结合用。对四批ICR纯种小鼠测得的K_D值为0.0918±0.0136nM。实验中用~3H-QNB测得结合曲线,用Scatchard、Hill及双倒数作图法求得K_D和B_~(max)值,还测定了非标记QNB取代50％时剂量、~5H-QNB的结合速度常数与解离速度常数及其t_(1/2)等参数。
In this paper, ~ 3H-QNB was used as specific binding ligand, and M receptor was detected by simple and sensitive radioligand method. The membrane preparation was prepared by low temperature centrifugation at 28000 × g, containing 0.8 mg of membrane protein in 2 ml of Na-K phosphate buffer (pH7.4) and incubated at 37 ° C. for half an hour with different concentrations of ~ 3H-QNB. The bound and free portions were assayed for nonspecific binding with 0.01 μM QNB. The KD value measured for four batches of ICR purebred mice was 0.0918 ± 0.0136 nM. The binding curve was measured with ~ 3H-QNB in the experiment. The K_D and B_max values were calculated by Scatchard, Hill and double reciprocal mapping. The dose of non-labeled QNB instead of 50% The combination of velocity constants and dissociation rate constants and their t_ (1/2) and other parameters.