Human il 1β elisa kit

THP1-derived M1 macrophages were treated as follows: (i) 1 × 10⁶ macrophages cultured alone; vi) 1 × 10⁶ macrophages cocultured with 2 × 10⁶ NSCs for 24 hours; (iii) 1 × 10⁶ macrophages cocultured with 2 × 10⁶ GSCs for 24 hours; (iv) 1 × 10⁶ macrophages noncontact cocultured with 2 × 10⁶ GSCs for 24 hours by 0.4 μmol/L Transwell, with GSCs placed in the upper chamber and BMDMs placed in the lower chamber; (v) 1 × 10⁶ macrophages pretreated with 1 μmol/L cytochalasin D for 1 hour at 37°C and then cocultured with 2 × 10⁶ U251 for 24 hours. ELISA and flow cytometry were performed on these cells.
1 × 10⁶ THP1-derived M1 macrophages were cocultured with 2 × 10⁶ GFP⁺ U251 for 24 hours, and then cells were stained with Pacific Blue anti-human CD45 (BioLegend, #982306, RRID:AB_2650649). CD45⁺GFP⁺ and CD45⁺GFP^– cells were sorted by fluorescence-activated cell sorting (FACS) for ELISA and flow cytometry analysis.
The protein levels of IL1α, IL1β, TNFα, and IL10 in the macrophage culture supernatant were measured with Human IL1α ELISA Kit (Cusabio, #CSB-E04620h), Human IL1β ELISA Kit (Cusabio, #CSB-E08053h), Human TNFα ELISA Kit (Cusabio, #CSB-E04740h), and Human IL10 ELISA Kit (Cusabio, #CSB-E04593h), respectively. Experiments were conducted according to the manufacturer's instructions.