Mouse elisa kit

The levels of nasopharyngeal cytokines and serum IgG of infected mice were determined using mouse ELISA kits (Sigma), with the exception of IL-17, which was performed using an enzyme-linked immunosorbent assay (ELISA) kit from eBiosciences. All ELISAs were performed according to the instructions of the manufacturer.

Blood was collected from cardiac puncture for blood chemistry and adipokine assays.
The enzyme activities of GOT and GPT, in addition to TCHO and TG levels were analyzed from serum samples using Fuji biochemical slides and a Fuji Dri-Chem 4000i analyzer (Fujifilm Cooperation, Taipei, Taiwan) in the Taiwan Mouse Clinic at Academia Sinica. Plasma levels of adiponectin, leptin, IL-6, and insulin were measured using mouse ELISA kits (Merck Millipore, Taipei, Taiwan). The plasma free fatty acid was measured using an ELISA kit (ab65341, Abcam, Cambridge, MA, USA), and blood HbA1c was measured using the mouse Hemoglobin A1c (HbA1c) Assay kit (Crystal Chem, Elk Grove Village, IL, USA).

For blood chemistry and adipokine assays, blood was collected from cardiac puncture at the conclusion of experiments. The TCHO levels were analyzed from serum samples using Fuji biochemical slides and a Fuji Dri-Chem 4000i analyzer (Fujifilm Cooperation, Taipei, Taiwan) in the Taiwan Mouse Clinic at Academia Sinica. Plasma levels of adiponectin, leptin, IL-6, and insulin were measured using mouse ELISA kits (Merck Millipore, Taipei, Taiwan). The plasma free fatty acids were measured using an ELISA kit (ab65341, Abcam, Cambridge, MA, USA), and blood HbA1c was measured using the mouse Hemoglobin A1c (HbA1c) Assay Kit (Crystal Chem, Elk Grove Village, IL, USA).

C5a (10⁻⁸ M) was added to untreated macrophages or those that had been pretreated with C4a (10⁻⁸ M) for 10 min. Then, the supernatant was collected for analysis of TNF-α, IL-6 and MCP-1 by using the mouse ELISA kit (Sigma). The ELISA plates were coated with 100 μl capture antibody per well at 4°C overnight. Following an appropriate wash, 200 μl of assay dilution buffer was added per well for inhibition at room temperature for 1 h. The sample and serial dilutions of the standards were added to the plate and incubated at 4°C overnight. Following coating with detection antibody, avidin-HRP was added and incubated at room temperature for 30 min. The substrate 3,3′,5,5′-tetramethylbenzidine was added and incubated for 15 min. Finally, 2N H₂SO₄ was added to terminate the reaction and the absorbance at 450 nm was measured using an ELISA reader (MTP-800 Microplate reader; Corona Electric, Tokyo, Japan).