Human il 1β elisa kit

For this assay, cells were exposed for 11 days with Hg(II) or MeHg as described in section 2.4. After exposure, apical and basolateral media were recovered for analysis of the pro-inflammatory cytokines IL-8 and IL-1β using Human IL-8/CXCL8 ELISA kit (Sigma) and Human IL-1β ELISA kit (Sigma), following the manufacturer instructions. To determine the influence of macrophages in the pro-inflammatory response of intestinal cells, a parallel exposure assay with Hg(II) or MeHg was performed using monolayers of Caco-2/HT29-MTX without the incorporation of differentiated THP-1 cells into the basolateral compartment. The conditions of seeding and Hg treatment were identical to those described above. The identification of the inflammatory pathways triggered by Hg was carried out using inhibitors. Cells were seeded as described above, Hg treatments were applied to the apical side, while inhibitors were added to the basolateral medium during the duration of the exposure, changing media every 2 days. The inhibitors used were: SP600125 (300 nM), BIRB796 (300 nM), BAY 11-7082 (250 nM) (MedChemExpress®). After exposure (6 days), cells were collected to analyse the gene