V plex mouse cytokine 19 plex kit

Inflammatory cytokines (TNF-alpha, IL-6 and IL-10) were quantified in serum, peritoneal fluid and organs collected in mice following CLP and in control mice, and supernatants from in vitro LPS-stimulated peritoneal macrophages. Uncoated ELISA kits were purchased from Invitrogen (Thermo Fisher Scientific) and ELISA were performed following manufacturer’s protocol: TNF-alpha (Invitrogen, #88-7324-22), IL-6 (Invitrogen, #88-7064-22), IL-10 (Invitrogen, #88-7105-22). Plates were red using Tecan.
Serum cytokines were measured using electrochemiluminescence assays (Meso Scale Discovery (MSD), Rockville, Maryland, USA). Serum TNF-alpha, IL-6, IL-10 and IL-33 were measured using a multiplex assay using the V-PLEX Mouse Cytokine 19-Plex kit (MSD). Samples were diluted 1:3 in proprietary buffer (MSD) and measured in duplicate according to manufacture's protocol. Plates were run on MSD plate reader model 1250.

Immunocompetent 8-week-old C57BL/6J male mice (Charles River Laboratories) were i.v. infected with a single dose (1 × 10¹¹ VP diluted in 100 μl PBS) of HAdV-20-42-42-LacZ (n = 8) or HAdV-5-LacZ (n = 8). Matched control groups were injected with 100 μl of PBS (n = 4). At 6 h after virus delivery, animals were sacrificed and blood was collected through cardiac puncture for serum isolation and subsequent cytokine analysis. Cytokine profiles were determined from 6 h postinjection sera using a V-PLEX mouse cytokine 19-plex kit (MSD) and a MESO SECTOR s 600 machine (Meso Scale Discovery).