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. Author manuscript; available in PMC: 2021 Apr 14.
Published in final edited form as: J Vis Exp. 2020 Apr 10;(158):10.3791/60517. doi: 10.3791/60517

Figure 4. Representative results for the LPS and LOS extraction and visualization procedure to test inner membrane (IM) purity.

Figure 4.

The volume of membrane sample corresponding to 50 μg of total protein was used to extract LPS from the IM and outer membrane (OM) of S. Typhimurium and E. coli DH5-alpha. The volume of membrane sample corresponding to 300 μg of total protein was used to extract LPS from the membranes of A. baumannii. The volumes were normalized to 100 μl with endotoxin-free water and treated with Proteinase K. LPS or LOS were extracted by hot-phenol extraction (1:1 water:phenol) and 21 μl of the extracts were loaded onto a 4–20% gradient polyacrylamide gel and visualized by PRO-Q Emerald 300 staining to assess the cross contamination of IM fractions with OM materials.