Smith T, Sutherland F, Chisholm G D, Habib F K
Clin Chim Acta. 1983 Jun 30;131(1-2):129-41. doi: 10.1016/0009-8981(83)90359-5.
Several factors affecting the reproducibility of androgen receptor assays in the human prostate gland have been identified. Experiments in the presence of proteolytic inhibitors such as PMSF, leupeptin and aprotinin resulted in no change in the estimates of androgen receptors. The androgen binding was also unaffected by the concentrations of endogenous steroids in the tissue. Although the receptor assay was reliable down to protein concentrations of about 5 g/l, at lower concentrations the reliability of the method was significantly reduced. Similarly, tissue storage at -25 degrees C drastically lowered the binding capacity of the prostatic tissue extracts to 50% of initial receptor content over a period of 3 months. The high degree of correlation between receptor levels in the stroma and epithelium suggests that the binding in the total gland will not be drastically affected by the proportion of stroma and epithelium it contains. Following the widely practised 2-h extraction of the nucleus with 0.6 mol/l KCl buffer, a highly significant non-extractable binding component remained. Analysis of data by the single-point assay produced results comparable to those obtained by standard Scatchard analysis and seemed reliable so long as careful quality control was employed.
已确定了几个影响人前列腺中雄激素受体检测重现性的因素。在存在蛋白酶抑制剂如苯甲基磺酰氟（PMSF）、亮抑酶肽和抑肽酶的情况下进行实验，雄激素受体的估计值没有变化。雄激素结合也不受组织中内源性类固醇浓度的影响。尽管受体检测在蛋白质浓度低至约5g/l时是可靠的，但在较低浓度下，该方法的可靠性显著降低。同样，在-25℃下储存组织，在3个月的时间内，前列腺组织提取物的结合能力急剧下降至初始受体含量的50%。基质和上皮中受体水平的高度相关性表明，腺体中所含基质和上皮的比例不会对全腺中的结合产生重大影响。在用0.6mol/l KCl缓冲液对细胞核进行广泛采用的2小时提取后，仍保留了一个高度显著的不可提取结合成分。通过单点检测分析数据产生的结果与通过标准Scatchard分析获得的结果相当，并且只要采用仔细的质量控制似乎就是可靠的。
