Le Fevre B, Bailly A, Sallas N, Milgrom E
Biochim Biophys Acta. 1979 Jun 12;585(2):266-72. doi: 10.1016/0304-4165(79)90026-6.
When soluble steroid-receptor complexes are exposed to DNA-cellulose only activated complexes bind. The specificity of the binding was shown by its dependence on the presence of hormone during activation. However, prolonged incubation of non-activated steroid-receptor complexes with DNA-cellulose led to a progressive activation of these complexes. When the same hepatic cytosol containing heat-activated [3H]triamcinolone acetonide-receptor complexes was titrated by high concentrations of nuclei or DNA-cellulose the former bound 75% of the complexes, the later only 40%. This decreased binding was due on the one hand to a lower initial interaction between DNA-cellulose and activated complexes than between nuclei and these complexes and on the other hand to increased losses during washes when DNA-cellulose was used. For these reasons nuclei and not DNA-cellulose should be used when accurate measurements of the concentration of activated complexes are required. When only comparative data are needed DNA-cellulose may, however, be employed.
当可溶性类固醇 - 受体复合物与DNA - 纤维素接触时，只有活化的复合物会结合。结合的特异性通过其在活化过程中对激素存在的依赖性得以体现。然而，未活化的类固醇 - 受体复合物与DNA - 纤维素长时间孵育会导致这些复合物逐渐活化。当用高浓度的细胞核或DNA - 纤维素滴定含有热活化的[³H]曲安奈德 - 受体复合物的相同肝细胞溶质时，前者结合了75%的复合物，后者仅结合40%。这种结合减少一方面是由于DNA - 纤维素与活化复合物之间的初始相互作用低于细胞核与这些复合物之间的相互作用，另一方面是由于使用DNA - 纤维素时洗涤过程中的损失增加。由于这些原因，当需要精确测量活化复合物的浓度时，应使用细胞核而非DNA - 纤维素。然而，当只需要比较数据时，可以使用DNA - 纤维素。
