Mouse ifn alpha elisa kit

Wildtype mice were treated intranasally with saline (40 µl) or 5′-ASO#26 (400 µg in 40 µl saline) once-daily for three days. Mice treated intranasally with lipopolysaccharides (60 µg in 40 µl saline, LPS from Escherichia coli O111:B4, Sigma) for one day were used as positive control. One day after the last administration, mice were sacrificed under anesthesia with 3% isoflurane, then BALF samples were collected by making an incision in the trachea and washing the lungs 5 times with 1 mL PBS, and repeating again with additional 1 ml PBS. BALF samples from each mouse were centrifuged at 450 g for 5 min at 4 °C and the supernatant was used for cytokine/chemokine measurement using the Bio-Plex MAGPix multiplex reader and Bio-Plex Pro Mouse Chemokine Panel 31-Plex panel (BioRad) following the manufacturer’s instructions. In addition, IFN alpha in BALF was measured with a Mouse IFN-alpha ELISA Kit (R&D system) following the manufacturer’s instructions.

Tumor necrosis factor (TNF)-α (Mouse TNF-alpha Quantikine enzyme-linked immunosorbent assay (ELISA) Kit, sensitivity: 7.2 pg/ml), interferon (IFN)-α (Mouse IFN-alpha ELISA Kit, sensitivity: 12.5 pg/ml), IFN-β (Mouse IFN-beta ELISA Kit, sensitivity: 15.5 pg/ml), IFN-γ (Mouse IFN-gamma Quantikine ELISA Kit, sensitivity: 2 pg/ml), interleukin (IL)-6 (Mouse IL-6 Quantikine ELISA Kit, sensitivity: 1.8 pg/ml), IL-8 (Mouse IL-8 Quantikine ELISA Kit, sensitivity: 2 pg/ml), IL-10 (Mouse IL-10 Quantikine ELISA Kit, sensitivity: 5.2 pg/ml), and monocyte chemoattractant protein (MCP)-1 (Mouse/Rat CCL2/JE/MCP-1 Quantikine ELISA Kit, sensitivity: 2 pg/ml) concentrations in serum and BAL were measured with commercially available ELISA technique kits following the manufacturer’s recommendations (R&D Systems, MN, USA).

Mouse IL‐12p70 Quantikine ELISA Kit (M1270), Mouse IL‐10 Quantikine ELISA Kit (M1000B), Mouse IFN‐beta Quantikine ELISA Kit (MIFNB0) and Mouse IFN‐alpha ELISA Kit (42120‐1), Human IL‐10 Quantikine ELISA Kit (D1000B), Human IL‐12 p70 Quantikine ELISA Kit (D1200) were purchased from R&D Systems. Mouse IL‐6 ELISA Kit (70‐EK206/3‐96) and Mouse TNF‐a ELISA Kit (70‐EK282/3‐96) were purchased from Multisciences (Lianke) Biotech, Co., Ltd. The methods were performed according to the manufacturer's instructions.