Mouse elisa kit

The endotoxemia model was employed as previously reported (41 (link), 56 (link)). Briefly, E. coli O26:B6 LPS (Sigma-Aldrich) was administered intranasally to induce endotoxemia (18 mg/kg). PBS and 5 mg/kg gcIFN-20 were intraperitoneally injected at 1 h after LPS treatment, respectively. Blood was collected via the tail vein at 6 h post LPS treatment. Whole blood was centrifuged at 3000 rpm at 4°C for 10 min, and the supernatant was collected. Serum TNF-α and IL-6 were measured using mouse ELISA kits (Multisciences). At 12 h after LPS treatment, all the mice were sacrificed, and lung tissues were collected. The pathological changes in lungs were evaluated after the slices were stained with HE, and NF-κB p65 protein was analyzed by immunohistochemistry.

RAW 264.7 cells were cultured in 24-well plates with 3 × 10⁵ cells per well for 24 h at 37°C under 5% CO₂. The cells were washed three times with PBS, and 300 μL of Dulbecco's Modified Eagle Medium (DMEM) containing LPS (E. coli O26:B6 LPS [Sigma-Aldrich], 150 ng/mL) and gcIFN-20 (0–128 μg/mL) was added to each well. Dexamethasone (1 μM) serves as the positive control. After incubation overnight, the supernatant was harvested by centrifuging the culture at 1500 g for 10 min at 4°C. The concentration of NO was determined by a nitric oxide assay kit (Beyotime Biotechnology), while the concentration of TNF-α was examined by mouse ELISA kits (Multisciences).

Blood samples were centrifuged at 7000 rpm for 7 min, at 4°C, and then the supernatants were collected. Colonic tissues were weighed, homogenized, and centrifuged at 12,000 rpm for 15 min, at 4°C, and the supernatants were collected. The cytokine levels of the cytokines IL-10, IL-6, and TNF-α were quantified using a corresponding mouse ELISA kits (MULTISCIENCES, China) according to the manufacturer’s instructions.

RAW 264.7 cells were cultured in 24-well plates with 3 × 10⁵ cells per well for 24 h at 37°C under 5% CO₂. The cells were washed three times with PBS, and 300 μL of Dulbecco's Modified Eagle Medium (DMEM) containing LPS (E. coli O26:B6 LPS [Sigma-Aldrich], 150 ng/mL) and gcIFN-20 (0–128 μg/mL) was added to each well. Dexamethasone (1 μM) serves as the positive control. After incubation overnight, the supernatant was harvested by centrifuging the culture at 1500 g for 10 min at 4°C. The concentration of NO was determined by a nitric oxide assay kit (Beyotime Biotechnology), while the concentration of TNF-α was examined by mouse ELISA kits (Multisciences).