Svec F, Tate D
Department of Medicine, Louisiana State University Medical Center, New Orleans 70112.
J Recept Res. 1987;7(6):859-70. doi: 10.3109/10799898709054566.
This study analyzes the effect of including molybdate in the elution buffers used in DEAE cellulose chromatography on the fraction of glucocorticoid receptor which elutes as the transformed species. Inclusion of molybdate leads to a significant decrease in the fraction of receptor eluting as transformed; samples which appear to be nearly 50% transformed if eluted in the absence of molybdate were found to be less than 10% transformed when analyzed using a buffer which contained 5 mM molybdate. This decrease was not caused by loss of receptor or a reversion of transformation. DEAE cellulose accelerates receptor transformation. It is concluded that DEAE cellulose should not be used to quantitate transformation unless molybdate is included in all buffers.
本研究分析了在DEAE纤维素色谱洗脱缓冲液中加入钼酸盐对以转化形式洗脱的糖皮质激素受体部分的影响。加入钼酸盐会导致以转化形式洗脱的受体部分显著减少；如果在不含钼酸盐的情况下洗脱，似乎近50%转化的样品，在用含有5 mM钼酸盐的缓冲液分析时，发现转化程度小于10%。这种减少不是由受体丢失或转化逆转引起的。DEAE纤维素会加速受体转化。得出的结论是，除非在所有缓冲液中都加入钼酸盐，否则不应使用DEAE纤维素来定量转化。
