R Lauro, R De Pirro, R Longhi, A Fusco, S Iacobelli
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引用次数: 0
摘要
采用密度梯度离心法研究大鼠肾受体对地塞米松的作用。用3h -地塞米松在Tris 10 mM, EDTA 1.5 mM,甘油5%,单硫甘油12 mM (pH 7.5)缓冲液中培养的细胞质进行密度梯度,由于复合物的不稳定性,从未给出任何85放射性峰的良好证据。通过将组织匀浆并将细胞质液在蒸馏水,甘油5%单硫甘油酯12mm溶液中孵育,达到复合物至少30小时的稳定性。密度梯度离心分析表明,放射性峰值出现在管的底部,可能是由于配合物的聚集。由于这种缺点是技术固有的,因此必须得出结论,密度梯度离心分析不能用于研究肾脏的地塞米松受体复合物。
[Attempts to characterize dexamethasone receptors present in rat kidney cytosol].
Density gradient centrifugation analysis has been used to study rat kidney receptors for dexamethasone. Density gradients performed with cytosol incubated with 3H-dexamethasone in a Tris 10 mM, EDTA 1.5 mM, glycerol 5%, monothioglycerol 12 mM (pH 7.5) buffer never gave any good evidence of a 85 radioactivity peak probably due to instability of the complex. The stability of the complex for at least 30 h was achieved by homogenizing the tissue and incubating cytosol in a distilled water, glycerol 5% monothioglycerol 12 mM solution. Density gradient centrifugation analysis in this way showed radioactivity peak at the bottom of the tube probably due to the aggregation of the complex. Since this drawback is inherent in the technique it must be concluded that density gradient centrifugation analysis cannot be used to study the dexamethasone-receptor complex of the kidney.