This work is supported by a grant from National Natural Sciences Foundation of China (39330210).
用NaOH代替苯乙胺作为14CO2的吸附剂,改进谷氨酸脱羧酶(GAD)活性的放射测定方法,结果发现NaOH为吸附剂组内变异系数为9.6%, 以苯乙胺为吸附剂组内变异系数为31.9%;以NaOH为吸附剂72 h后测量其放射活性仍稳定不变,以苯乙胺为吸附剂者1 h后放射性活性即下降47%,6 h后已降低至本底水平;14CO2重吸收实验亦证明以苯乙胺为吸附剂吸附的14CO2 6 h内已有80%以上重新被NaOH吸附;以NaOH作为吸附剂测定GAD的活性,在0.39~17.8 mg脑组织样品范围内GAD量与14CO2生成量之间有线性关系.NaOH代替苯乙胺作为14CO2的吸附剂测定GAD的活性其灵敏度提高1.66倍.用此方法测定组织和细胞内GAD活性证明其具有良好的重复性和稳定性,值得推广应用.
The radioassay of glutamic decarboxylase(GAD) was modified by taking NaOH as trapped agent instead of phenylethylamine. The results showed that the coefficient of variation (CV) within same sample was 9.6% and the radioactivity remains stable after 72 hours if use NaOH as trap agent. It is significantly stable than use phenylethylamine as trap agent, which the CV was 31.9% and the radioactivity decreased 47% within the first hour and decreased to background after 6 hours. The reabsorption experiment shows over 80% of 14CO2 can be reabsorption by NaOH within 6 hours. It is suggested that NaOH is a much better trap agent than phenylethylamine and the sensitivity can increase 1.66 folds. Using this method the GAD activity in 0.39~17.8 mg of brain tissue can be measured and it is success in determine the GAD activity both in rat brain tissue and cultured neurons.
胡元元,何善述.谷氨酸脱羧酶放射测量法的改良及应用[J].生物化学与生物物理进展,2001,28(1):118-120
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