Quorum Sensing
Chapter 4 Liquid Chromatography/Mass Spectrometry (LC/MS) for the Detection and Quantification ofN-Acyl-L-Homoserine Lactones (A ...
2 Materials 2.1 Bacterial Cell Cultures Bacterial strain: The methods presented here use cultures of the Gram-negative bacteriu ...
unit (14 Da) going from one congener to the next. The most abundant HAQs, contrary to AHLs, contain an odd number of carbons on ...
or other metabolites directly in culture broth. In multiple reaction monitoring (MRM) mode, one takes advantages of the ability ...
Fig. 2Total ion chromatogram of the supernatant of aP. aeruginosastrain PA14 culture grown to an OD 600 of 3.0 Fig. 3Ion chromat ...
The area under each of these chromatographic peaks is integrated. The concentration of the analyte A in the culture medium is g ...
The following transitions are monitored: C 4 -HSL 172!102; o-C 4 -HSL 190!120; HHQ-d 4248 !163; 3-oxo-C 12 -HSL 298 !102; and o ...
Using a Pasteur pipette, the organic phase (top when ethyl acetate is used) is transferred to a HPLC vial, being very careful n ...
normal scanning mode. However, HHQ and the other mem- bers of the same family of compounds are the precursors of PQS and its con ...
Because the Las system, which mediates 3-oxo-C 12 -HSL, is expressed before the Rhl system which produces the C 4 -HSL, the max ...
position that is required for quorum-sensing regulation. J Bacteriol 190:5339–5352 Rahme LG, Stevens EJ, Wolfort SF, Shao J, To ...
Chapter 5 Detection of the Bacterial Quorum-Sensing Signaling MoleculesN-Acyl-Homoserine Lactones (HSL) andN-Acyl-Homoserine (HS ...
molecules have been shown to be relevant in natural systems. Furthermore, it is often not possible to detect the degradation pro ...
2 Materials All chemicals should be at least of analytical grade and LC-MS- grade solvents needed for chromatographic separation ...
Hydrolysis: 1 M NaOH, 1 M HCl. Substrate: 0.4 mM 1-Step Ultra Tetramethylbenzidine (TMB)-ELISA, 1.3 mM H 2 O 2 in 100 mM sodium ...
2.3 HSL/HS Extraction of Cell Pellets Extraction solvent: Prepare 100 ml of anhydrous ethyl acetate and acidify the solution wi ...
General washing procedure: Wash the coated plate three times with 200μl of 4 mM PBST buffer per well. This can be done manually ...
Evaluation: Curve fitting of the standard curve can be performed using the four-parameter fit according to the equa- tion [10] ...
c(HSL) andc(HS) are the individual concentrations of HSL and HS in the samples, whileS(HSL) hydrolyzed and non- hydrolyzed and S ...
rate of 0.1 ml/min. Change solvent composition after 20 min first to 50% eluant B and second to 0.5% eluant B. Set sample tempe ...
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