Ni ida 2000 packed columns protino

Plasmids of mouse vimentins provided by Prof. Dr. Yi-Ling Li, Institute of Biomedical Sciences, Genomics Research Center, Academia Sinica, Taipei, Taiwan, were used (Liang et al.55 (link)). The plasmids were transformed into BL21(DE3) cells. Induction was with 1 mm isopropyl 1-thio-β-d-galactopyranoside at 37 °C for 3 h. Recombinant vimentin proteins were purified as described by Machery-Nagel for His tag protein purification (Ni-IDA 2000 Packed Columns Protino, Machery-Nagel GmbH and Co. KG, Düren, Germany). Eluted proteins were analyzed by 15% SDS-PAGE, stained with Coomassie Blue, and submitted to Western blot analysis against α-penta-His (catalog number 34660, Qiagen, Hilden, Germany).

Plasmids of mouse vimentins provided by Prof. Dr. Yi-Ling Li, Institute of Biomedical Sciences, Genomics Research Center, Academia Sinica, Taipei, Taiwan were used [25] (link). The plasmids were transformed into BL21 (DE3) cells. Induction was with 1 mM IPTG at 37°C for 3 h. Recombinant vimentin proteins were purified as described by Machery-Nagel for His-tag protein purification (Ni-IDA 2000 Packed Columns Protino, Machery-Nagel GmbH & Co. KG, Düren, Germany). Eluted proteins were analyzed by 15% SDS-PAGE, stained with Coomassie blue and submitted to Western blot analysis against α-penta-His (Qiagen (catalog no. 34660), Hilden, Germany).