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开发了两步亲和色谱法:肝素-琼脂糖凝胶、Ni-琼脂糖凝胶色谱纯化人血浆中硒蛋白-P的方法,并采用氢化物发生-原子荧光分光光度法(HG-AFS)检测,成功搭建了硒蛋白-P的纯化检测平台。确定了亲和色谱纯化的最佳梯度洗脱条件,通过十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)定性检测,得到了一定纯度的硒蛋白-P,其回收率达43.2%。HG-AFS方法的线性相关系数为0.999 1,检出限为0.09μg/L,日内精密度(RSD)为0.12%,日间精密度(RSD)为0.27%,加标回收率为95%~104%。该亲和色谱纯化方法简单易控、回收率高,HG-AFS检测灵敏度高,结果准确可靠。 相似文献
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Heparin-sepharose forms complexes with putrescine, spermidine, and spermine and indirect measurements of its affinity for
polyamines gives values similar to those obtained with free heparin. A direct measurement of the binding of heparin-sepharose
to spermine gives an apparent dissociation constant (K
d) of 1.5×10−6
M spermine.
Unlike free heparin, heparin-sepharose does not cause either disruption of the nuclei or more sutble modifications able to
modify their sedimentation behavior.
The heparin-sepharose polyamine complex formed by the addition of heparin-sepharose to the homogenate can easily be removed
and the homogenate can be processed according to normal schedules.
Heparin-sepharose is able to sequester 85% of the exchangeable spermine present in the homogenate of rat ventral prostate.
The distribution of the marker enzyme galactosyltransferase (Golgi apparatus) on a sucrose density gradient was followed to
assess the usefulness of heparin-sepharose in minimizing the aggregation of cellular organelles brought about by polyamines. 相似文献
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