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_83
Snippet: Immunofluorescence microscopy: Immunofluorescence microscopy was performed as described previously (Buske and Levin, 2012) . Cells were grown using same media conditions overnight in 0.5% xylose, back-diluted 1:100 and grown in 0.5% xylose until the cells reached mid-log phase. The cells were then washed twice with LB, diluted 1:100, and grown in 0.1 mM IPTG for 5 generations (~2.5 hours). The cells were harvested and fixed with 16% paraformaldeh.....
Document: Immunofluorescence microscopy: Immunofluorescence microscopy was performed as described previously (Buske and Levin, 2012) . Cells were grown using same media conditions overnight in 0.5% xylose, back-diluted 1:100 and grown in 0.5% xylose until the cells reached mid-log phase. The cells were then washed twice with LB, diluted 1:100, and grown in 0.1 mM IPTG for 5 generations (~2.5 hours). The cells were harvested and fixed with 16% paraformaldehyde/0.5% gluteraldehyde. The cells were lysed with 2 mg/mL lysozyme. FtsZ was detected with affinity-purified polyclonal rabbit anti-FtsZ serum in combination with goat antirabbit serum conjugated to Alexa488 (Life Technologies). Cell walls were stained with wheatgerm agglutinin conjugated to tetramethylrhodamine, and DNA was stained with DAPI. All rights reserved. No reuse allowed without permission.
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