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Cytokine multi-plex ELISA

GO Grace C. O’Regan
SF Sahar H. Farag
GO Gary R. Ostroff
ST Sarah J. Tabrizi
RA Ralph Andre
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Cytokine profiling was carried out using MesoScale Discovery V-PLEX Proinflammatory Cytokine Panel 1 (Human) ELISA kits, following the manufacturer’s instructions. Supernatants from LPS-stimulated and unstimulated cultures were diluted 1:300 and 1:5, respectively. Three parallel wells as technical replicates were measured per single data-point. Data were normalised to total cell protein content as measured by Pierce BCA assay (Thermo Fisher Scientific), having been washed once with sterile D-PBS and lysed in 60 µl RIPA buffer supplemented with Roche cOmplete Mini EDTA-free protease inhibitor cocktail.

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