Mouse elisa kit

Hepatocyte cytokines, including IL-6 (DY406), TNF-α (DY410) and IL-1β (DY401), were measured using a commercial mouse ELISA kit (R&D Systems, Minneapolis, MN, USA), according to the manufacturer’s instructions.

Primary peritoneal macrophages were isolated from C57BL/6 mice using 3.85% thioglycollate solution, and treated with 400 μM PA and/or 100 ng/ml LPS. After incubation for 24 h, IL-1β content in culture supernatants was determined using a mouse ELISA kit (R&D Systems).

IFN-γ and granzyme B concentrations in the supernatants of CD8⁺ T cells were measured using a mouse ELISA kit (R&D Systems) following the manufacturer’s protocol. IFN-γ and granzyme B concentrations were within the range of the standard curve. All points were done in triplicate, and the experiments were repeated three times. Samples were read in a microplate reader (Infinite 200 PRO, Tecan).

The level of shh in the cell culture medium of astrocytes was measured by a mouse ELISA kit (R&D system, Minneapolis, MN, USA) according to the manufacturer’s instructions.

The mouse ELISA kit (R&D, Minneapolis, MN, Cat# DY479-05) was used for CCL2 assay. The blood was centrifuged at 1000×g, 4°C for 15 minutes and the serum was then collected for use. The ELISA plates were coated with 100 μl capture antibody at 4°C overnight. After wash, 200 μl of assay dilution buffer were added for blocking at room temperature for 1 hour. The samples and serial dilutions of standards were added and incubated at 4°C for overnight. After incubating with the detection antibody, avidin-HRP was added and incubated at room temperature for 30 minutes. The substrate, 3,3′,5,5′-tetramethylbenzidine (TMB), was then added and incubated for 15 minutes. 100 μl of stop solution were added to stop reaction, and the plate was then subjected to measurement of absorbance at 450 nm by using an ELISA reader.

Chemokines and cytokines were measured in supernatants of homogenized lung tissue using mouse TH1/TH2 9-PlexBase cytokine/chemokine multiplex assay (Meso Scale Discovery, Gaithersburg, Maryland, USA), IL-13 was measured using mouse ELISA Kit (R&D Systems, Minneapolis, USA), and total TGF-β 1 was measured with mouse TGF-ß1 Platinum ELISA (Bender MedSystems GmbH, Vienna, Austria), all according to manufactureŕs instructions.