Author: Megan C. Cohan; Ammon E. Posey; Steven J. Grigsby; Anuradha Mittal; Alex S. Holehouse; Paul J. Buske; Petra A. Levin; Rohit V. Pappu
Title: Evolved sequence features within the intrinsically disordered tail influence FtsZ assembly and bacterial cell division Document date: 2018_4_14
ID: 2rzfuy33_91
Snippet: The copyright holder for this preprint (which was not peer-reviewed) is the author/funder. . https://doi.org/10.1101/301622 doi: bioRxiv preprint for an additional 4h at 37ËšC, and then the cells were harvested by centrifugation, and the cell pellets were stored at -80ËšC. Purification was carried out as previously described (Buske and Levin, 2012) . Peak fractions were analyzed by SDS-PAGE and pooled together, and dialyzed overnight in 1 L of Ft.....
Document: The copyright holder for this preprint (which was not peer-reviewed) is the author/funder. . https://doi.org/10.1101/301622 doi: bioRxiv preprint for an additional 4h at 37ËšC, and then the cells were harvested by centrifugation, and the cell pellets were stored at -80ËšC. Purification was carried out as previously described (Buske and Levin, 2012) . Peak fractions were analyzed by SDS-PAGE and pooled together, and dialyzed overnight in 1 L of FtsZ dialysis buffer (50 mM MES 50 mM KCl, 2.5 mM MgCl 2 , 1 mM EGTA, 10% sucrose, pH 6.5) Protein preparations were concentrated, separated into aliquots, and flash frozen on liquid N 2 , and stored at -80ËšC. Prior to use, FtsZ aliquots were thawed on ice, well mixed, and the concentration was quantified using Pierce 660nm assay with tubulin as a standard (Thermo Fisher Scientific).
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