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doi: 10.1038/s41380-022-01479-w 279 SinghN.GuravA.SivaprakasamS.BradyE.PadiaR.ShiH.et al

S-glutathionylation regulates inflammatory activities of S100A9

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Materials for Detection of Product Formation from Live Cells by AlphaScreen Assays Medium-binding white solid-bottom tissue culture treated 1536-well plates Plate reader capable of 1536-well AlphaScreen detection (such as the EnVision Plate Reader equipped with a 1536 HTS AlphaScreen aperture) THP-1 cells in media with 5% serum ( see Note 4 ) Media with 5% serum Lipopolysaccharide (LPS) (4 g/mL working stock for 1 g/mL, final) in media with 5% serum Mixture of biotinylated anti-TNF- antibody (4.66 nM working stock for 1 nM, final) and anti-TNF--conjugated acceptor beads (46.66 g/mL working stock for 11 g/mL, final) in AlphaLISA immunoassay buffer Streptavidin donor beads (186.4 g/mL working stock for 40 g/mL, final) in AlphaLISA immunoassay buffer ( see Note 2 ) Experimental compounds suspended in DMSO (stock concentrations of 640 nM 10,000 M for a working concentration range of 3.7 nM 57.5 M after a 174-fold dilution into the assay mixture) Liquid transfer pin tool (such as a Kalypsys automated pin-tool station) ( see Note 3 ) Centrifuge equipped with a rotor that will accommodate plates Liquid dispenser capable of delivering small volumes (such as a Multidrop Combi Reagent Dispenser) 3

Many foods appear to act as both inducers and inhibitors of CYP1 enzymes, an effect which may be dose dependent or altered by the isolation of bioactive compounds derived from food