Elisa max deluxe set human il 1α

LDH was measured using an LDH Cytotoxicity Kit (Pierce, 88953), CyQUANT^™ LDH Cytotoxicity Assay (Invitrogen C2030), or Cytotoxicity Detection Kit (Sigma, 11644793001) per the manufacturer’s instructions; for any set of experiments, the same assay kit was used for all experiments. Measurements at 490 nm and 680 nm were taken immediately after the addition of Stop Solution using a BioTek Plate Reader. To determine LDH activity, the 680 nm absorbance value (background) was subtracted from the 490-nm absorbance value, and cytotoxicity was calculated for each sample as LDH activity as a fraction of the total LDH activity for cells treated with the Triton X-100 lysis buffer provided by the manufacturer. For experiments involving primary cells, a different donor was used for each biological replicate; due to inherent variability from donor to donor in the baseline levels of death in cell death assays, within-donor data were normalized to the negative control condition before compilation of the biological replicates. Assays of levels of secreted cytokines were performed using the Human Total IL-18 DuoSet ELISA (R&D Biosystems, DY318–05), ELISA MAX Deluxe Set Human IL-1α (Biolegend, 445804), and ELISA MAX Deluxe Set Human IL-1β (Biolegend, 437004), per the manufacturer’s instructions.