Copper Binding of Nuclear RNA- and DNA-Binding Proteins p54nrb/NonO and PSF
Copper Binding of Nuclear RNA- and DNA-Binding Proteins p54nrb/NonO and PSF
复制标题
核 RNA 和 DNA 结合蛋白 p54nrb/NonO 和 PSF 的铜结合
DOI：
10.11299/brte.17.328
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发表时间：
2006
期刊：
影响因子：
--
通讯作者：
M. Tezuka
中科院分区：
文献类型：
--
作者：
Masaki Watanabe;M. Tezuka
材料和方法Cu-IMAC和Western印迹。用FLAG标记的p54 nrb/NonO或PSF构建体转染人胚肾细胞系HEK 293。转染48 h后，用平衡缓冲液（20 mM磷酸钠缓冲液，pH 7.4，0.5 M NaCl）收获细胞，超声处理，然后以15，000 × g离心30 min。将上清液置于Cu-IMAC中。对于Cu-IMAC，将0.2mL HiTrap螯合HP（GE
Materials and Methods Cu-IMAC and Western blotting. The human embryonic kidney cell line HEK293 were transfected with FLAG-tagged p54nrb/NonO or PSF constructs. After 48 h of transfection, cells were harvested with equilibration buffer (20 mM sodium phosphate buffer, pH 7.4, 0.5 M NaCl), sonicated, and then centrifuged at 15,000× g for 30 min. The supernatants were subjected to Cu-IMAC. For Cu-IMAC, 0.2 mL of HiTrap Chelating HP (GE