Author: Jacob E. Choby; Hanna B. Buechi; Allison J. Farrand; Eric P. Skaar; Matthew F. Barber
Title: Molecular basis for the evolution of species-specific hemoglobin capture by pathogenic Staphylococcus Document date: 2018_6_7
ID: ieh5cvw1_47
Snippet: The copyright holder for this preprint (which was not peer-reviewed) is the author/funder. . https://doi.org/10.1101/339705 doi: bioRxiv preprint expression of hemoglobin was induced with 40 µg/ml IPTG (RPI, Mount Prospect, IL). After 16 h post-383 induction, cells were collected by centrifugation. The cell pellet was resuspended in PBS containing 10 384 mM imidazole (Fisher), 5 mM MgCl 2 (Sigma), 1 Roche Protease inhibitor table (Fisher), and a.....
Document: The copyright holder for this preprint (which was not peer-reviewed) is the author/funder. . https://doi.org/10.1101/339705 doi: bioRxiv preprint expression of hemoglobin was induced with 40 µg/ml IPTG (RPI, Mount Prospect, IL). After 16 h post-383 induction, cells were collected by centrifugation. The cell pellet was resuspended in PBS containing 10 384 mM imidazole (Fisher), 5 mM MgCl 2 (Sigma), 1 Roche Protease inhibitor table (Fisher), and a few mg of 385 lysozyme (Thermo) and deoxyribonuclease from bovine pancreas (Sigma). The cell pellet resuspended 386 with rocking for 20 min following incubation on ice. Cells were lysed using an Emulsiflex (Avestin, Ottawa, 387 CA) then cell lysate was clarified by ultracentrifugation (60 min at 17,000 g). Cell lysate was applied to a 388
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