Author: K. Reddisiva Prasanth; Minato Hirano; W. Samuel Fagg; Eileen T. McAnarney; Chao Shan; Xuping Xie; Adam Hage; Colette A. Pietzsch; Alexander Bukreyev; Ricardo Rajsbaum; Pei-Yong Shi; Mark T. Bedford; Shelton S. Bradrick; Vineet Menachery; Mariano A. Garcia-Blanco
Title: Topoisomerase III-ß is required for efficient replication of positive-sense RNA viruses Document date: 2020_3_27
ID: a0xfb9l4_21
Snippet: FLAG-TOP3 and crosslinked RNA complexes were immunoprecipitated from the 500 µl of total cell lysate by using Pierce TM Anti-DYKDDDK Magnetic Agarose beads (Thermo fisher scientific). After the extensive washing with high-salt buffer (1M NaCl 2 , 100mM Tris-HCl pH 7.4, 2.5m MgCl 2 , 0.5% NP-40, 0.5% Triton X-100, 0.1% SDS), the input cell-lysate, the depleted supernatant after IP and 25% of the beads were analyzed with Western-blotting and induc.....
Document: FLAG-TOP3 and crosslinked RNA complexes were immunoprecipitated from the 500 µl of total cell lysate by using Pierce TM Anti-DYKDDDK Magnetic Agarose beads (Thermo fisher scientific). After the extensive washing with high-salt buffer (1M NaCl 2 , 100mM Tris-HCl pH 7.4, 2.5m MgCl 2 , 0.5% NP-40, 0.5% Triton X-100, 0.1% SDS), the input cell-lysate, the depleted supernatant after IP and 25% of the beads were analyzed with Western-blotting and induced expression and IP were confirmed.
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