Mouse ifn α elisa kit

Brains at 8 day post infection were collected in PBS with 0.1 mg/ml soybean trypsin inhibitor (Sigma), 1.5 mM Pefabloc (Roche), 50 mM EDTA, and 0.1% bovine serum albumin and frozen at -20°C. After thawing, samples were permeabilized using 2% saponin over night at 4°C, and the supernatant was subsequently collected by centrifugation. Chemokine and cytokine analysis was performed using a mouse magnetic Luminex assay from R&D Systems for 23 cytokines/chemokines. Significant differences in comparison to control were detected for CCL2 [lowest level of quantification (LLOQ) = 51 pg/ml], CCL3 (LLOQ = 77 pg/ml), CCL4 (LLOQ = 75 pg/ml), CCL5 (LLOQ = 55 pg/ml), CXCL1 (LLOQ = 57 pg/ml), IFN-γ (LLOQ = 34 pg/ml), IL-6 (LLOQ = 38 pg/ml), IL-12 (LLOQ = 76 pg/ml), IL-13 (LLOQ = 37 pg/ml) according to manufacturer’s instructions and analyzed using Bio-Plex 200 system (Bio-Rad). The levels of CXCL9, CXCL10 and IFN-alpha were measured using Mouse CXCL9/MIG Quantikine ELISA Kit (R&D, Minneapolis, MN, USA, LLOQ 7.8 pg/ml), Mouse CXCL10/IP-10/CRG-2 DuoSet ELISA kit (R&D, LLOQ 62 pg/ml) and Mouse IFN-α ELISA Kit (R&D, LLOQ 12.5 pg/ml) according to manufacturer’s instruction.

The NK cell activity of mouse splenocytes was assessed using flow cytometry [22 (link)] as described previously [14 , 17 (link), 23 (link)]. The level of target cell lysis was determined using a Cytomics FC500 flow cytometer (Beckman Coulter, Brea, CA, USA), and the NK cell activity was expressed as the percentage of effector cell-specific lysis. Concentrations of IFN-α in BALF and nasal wash were determined with mouse IFN-α ELISA kit (R&D Systems, Inc., Minneapolis, MN, USA) according to manufacturer's instructions.