Details on the strains are given in Table 1. For LPS purification, a 10-microliter loop full of bacterial colony material was re-suspended thoroughly in 6 ml saline (0.9% NaCl, pH 7.0) up to a density comparable to McFarland scale 4.0. A volume of 3 ml of 37% formaldehyde was added to achieve a final formaldehyde concentration of 12.3%. The suspension was rigorously vortexed and subsequently swayed using an orbital shaker (Polymax 1040, Heidolph Instruments GmbH, Schwabach, Germany) over night at room temperature. Controls for sterility confirmed inactivation of
bacteria. Cells were pelletized by centrifugation at 3.500× g for 15 min and the supernatant was discarded. Soluble components were removed by 3 consecutive centri-
fugation and washing steps with 9 ml phosphate
buffered saline (PBS, pH 7.0) shaking at 15 rpm for 10
minutes. The LPS-containing pellet was re-suspended in
Details on the strains are given in Table 1. For LPS purification, a 10-microliter loop full of bacterial colony material was re-suspended thoroughly in 6 ml saline (0.9% NaCl, pH 7.0) up to a density comparable to McFarland scale 4.0. A volume of 3 ml of 37% formaldehyde was added to achieve a final formaldehyde concentration of 12.3%. The suspension was rigorously vortexed and subsequently swayed using an orbital shaker (Polymax 1040, Heidolph Instruments GmbH, Schwabach, Germany) over night at room temperature. Controls for sterility confirmed inactivation ofbacteria. Cells were pelletized by centrifugation at 3.500× g for 15 min and the supernatant was discarded. Soluble components were removed by 3 consecutive centri-fugation and washing steps with 9 ml phosphatebuffered saline (PBS, pH 7.0) shaking at 15 rpm for 10minutes. The LPS-containing pellet was re-suspended in
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