Lauro R, De Pirro R, Longhi R, Fusco A, Iacobelli S
Arch Sci Biol (Bologna). 1978 Jan-Dec;62(1-4):13-9.
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.
密度梯度离心分析已被用于研究大鼠肾脏中的地塞米松受体。在含有10 mM Tris、1.5 mM EDTA、5%甘油、12 mM单硫代甘油(pH 7.5)的缓冲液中,用3H-地塞米松孵育的细胞溶质进行密度梯度实验,从未得到任何关于可能由于复合物不稳定而出现的85放射性峰的良好证据。通过将组织匀浆并在含有5%甘油、12 mM单硫代甘油的蒸馏水溶液中孵育细胞溶质,可使复合物的稳定性至少达到30小时。以这种方式进行密度梯度离心分析时,在管底部出现了放射性峰,这可能是由于复合物的聚集所致。由于该技术存在这一固有缺陷,因此必须得出结论,密度梯度离心分析不能用于研究肾脏中的地塞米松-受体复合物。