Max deluxe set human

THP-1 cells (9 × 10⁴) were seeded in flat-bottomed 96-well plates with/without 200 ng/mL LPS (Enzo) and incubated for 2, 4, 6, 12, and 24 h. Following treatment, plates were centrifuged at 130 × g for 5 min at room temperature to pellet cells. Supernatants were transferred to new 96-well plates and stored at −80 °C. ELISA kits for IL-1β (BioLegend, Max Deluxe Set Human), TNFα (BioLegend, Max Deluxe Set Human), IL-6 (BioLegend, Max Deluxe Set Human), CXCL10 (R&D Systems, Human DuoSet), and IL-8 (R&D Systems, Human DuoSet) were performed following manufacturer’s instructions using Nunc MaxiSorp 96 well flat-bottomed ELISA plates (Thermo Fisher Scientific). Supernatants were prepared in the appropriate reagent diluent using the following dilution factor: IL-1β, neat; TNFα, 1:15; IL-6, 1:5; CXCL10, 1:5; and IL-8, 1:20. ELISA plates were read on a CLARIOstar microplate reader (BMG LABTECH), with cytokine concentrations extrapolated using the MARS Data Analysis Software (BMG LABTECH).