Opteia human il 1β elisa set

The BD OptEIA Mouse IL-1β ELISA Set and BD OptEIA Human IL-1β ELISA Set (BD Biosciences), Mouse IL-18 ELISA Kit (MBL International), TNF alpha Human ELISA Kit (abcam) and Mouse TNF-α DuoSet (R&D Systems) were used according to the manufacturers’ instructions. LDH release was determined by CytoTox-ONE Homogeneous Membrane Integrity Assay (Promega) according to the manufacturers’ instructions.

Human primary bone marrow mononuclear cells were purchased from ATCC, Manassas, VA, USA (catalog number PCS-800-013). The cells were plated onto either a 96-well plate (4 × 10⁴ cells/well) or a 12-well plate (5 × 10⁵ cells/well), and were differentiated to macrophages for seven days in RPMI1640 containing 10% FBS and 50 ng/mL M-CSF. The culture medium was then replaced with RPMI1640 containing 10% FBS and lipopolysaccharide (0.5 μg/mL). After 2 h of incubation, the cells were washed with RPMI-1640 and were pretreated with either DMSO, MCC950, or extracts for 1 h in RPMI-1640 without any supplement, and then were treated with either SiO₂ (100 μg/mL) or TiO₂ (100 μg/mL) nanoparticles for 3 h. The conditioned media were used for either ELISA using BD OptEIA Human IL-1β ELISA set (BD Bioscience, San Jose, CA, USA) or Western blot analysis.

Supernatants of monocyte derived macrophages were collected after stimulation and analyzed for TNF, IL1β and NO release. The BD OptEIA™ Human TNF ELISA Set(BD Biosciences, USA), BD OptEIA™ Human IL1β ELISA Set (BD Biosciences, USA), and Total Nitric Oxide and Nitrate/Nitrite Parameter Assay Kit (Bio-Techne, Germany) were applied according to the manufacturer’s protocol.