Elisa kit

IFNγ, IL-4 and IL-17 secretion from cell culture supernatants of iNKT cell and KMC functional assays were assessed by either a Milliplex_MAP Kit (Millipore) on a Bio-Plex 200 system (Bio-Rad) or Ready-Set-Go! ELISA kit (eBioscience). IL-21 secretions were analyzed by a Milliplex_MAP Kit (Millipore) on a Bio-Plex 200 system (Bio-Rad). Measurements of IgM and IgG in culture supernatants of B cell help assays were performed using an ELISA kit (SouthernBiotech) as described previously [19 (link)]. Anti-dsDNA IgG in the supernatants of whole splenocyte cultures and BW Ly108^hi/loiNKT and B cells co-cultures were assayed by mouse anti-dsDNA IgG-specific ELISA kits (Alpha Diagnostic Intl. Inc.). Anti-dsDNA IgG and other IgG autoantibodies from co-culture supernatants were assayed using nucleic acid/proteomic microarray plates described below.

Eight-week-old female BALB/c mice (n = 8 per group) were challenged intranasally with 8 μg/mouse ovalbumin (OVA, Grade V, A5503, Sigma-Aldrich, St. Louis, MO, USA) alone or in combination with 60 μL of 1 μM pH-balanced 8-oxoG solution on days 0–4, 18 and 28 under mild anesthesia. In controls, intranasal challenges were performed with 60 μL of PBS. On day 30, animals were sacrificed and sera were collected for the measurement of IgM or IgE production. The levels of OVA-specific IgM and IgE in the mice’ sera were measured by ELISA kit (Southern Biotech, Birmingham, AL, USA), according to the manufacturer’s instructions. Absorbance was detected by a Synergy HT microplate reader (Bio-Tek Instruments, Winooski, VT, USA) at 450 nm.