Human il 1β elisa kit

The conditioned medium of HMC3 cells was collected and centrifuged at 1000× g for 5 min. Then, the supernatant was passed through a 0.22 µm filter to eliminate smaller debris and was placed on dry ice for snap-freezing and stored at −80 °C until use. Human IL-1 β ELISA Kit (invitrogen), IL-18 ELISA Kit (invitrogen), and c-Myc ELISA Kit (invitrogen) were used according to manufacturer’s protocol. 50 µL of the conditioned medium was used in each well of ELISA plates to detect. The plates were then analyzed on a SpectraMax M2 Microplate Reader (Molecular Devices, Silicon Valley, CA, USA) and the absorbance was read at 450 nm.

THP-1
cells were exposed to uncoated or surface-modified (Al and ES) NPs
(2.5 μg/mL) for 6 h, and samples were collected and stored at
−80 °C. IL-1β release was determined using a human
IL-1β ELISA kit (Invitrogen, Sweden) according to the manufacturer’s
instruction. Absorbance was measured at 450 nm using a Tecan Infinite
F200 plate reader. Results are expressed as pg/50.000 cells of released
cytokine, based on at least three independent experiments. For experiments
using different inhibitors, cells were pre-incubated for 1 h with
the indicated inhibitors, z-VAD-fmk (20 μM, R&D Systems),
CaO74Me (10 μM, Sigma-Aldrich), A438079 (50 μM, Sigma-Aldrich),
Trolox (500 μM, Sigma-Aldrich), and MCC950 (10 μM, Sigma-Aldrich).
Then, cells were exposed to uncoated or surface-modified (Al and ES)
NPs, cell media were collected, and IL-1β quantification was
performed as described above. To validate the role of NLRP3, the Null-1
and defNLRP3 THP-1 cell lines were used.^{47 (link)} The cells were grown with selection antibiotics (see: “inflammasome
knockout cells”). Cells were plated at a density of 10⁵ cells/well and exposed for 6 h to bare and surface-modified
silica NPs (2.5 μg/mL) in medium supplemented with 10% FBS.
Cell media were collected, and IL-1β content was measured using
a specific ELISA. Additionally, TNF-α was determined using the
TNF alpha ELISA kit (Invitrogen, Sweden).

The cytokine production of monocytes was measured by ELISA. A total of 1 × 10⁵ cells of monocytes were added to each hCO of 6 weeks old with 100 µL of cortical organoid media III. After culturing in a 37 °C incubator for 24 h, 90 µL medium was aspirated into an Eppendorf tube and centrifuged at 300 g for 10 min. After centrifugation, 80 µL of the supernatant was diluted to 250 µL. Concentrations of MCP‐3, IL‐1β, and TNF‐α were measured by using Quantikine ELISA Human CCL7/MCP‐3 (R&D systems), Human IL‐1β ELISA Kit (Invitrogen), and Human TNF‐α ELISA Kit (Invitrogen), respectively. For each ELISA measurement, 50 µL of the diluted culture medium was used. The results were read by a Synergy H1 plate reader (BioTek). The standard curve was used to calculate the concentration of cytokines.

Cell supernatant was collected, and cytokine production was analyzed with Human TNF-α ELISA kit (Invitrogen, cat. no. KHC3011), Human IL-1β ELISA kit (Invitrogen, cat. no. BMS224–2), and Human IL-8 ELISA kit (Invitrogen, cat. no. BMS204–3), following the manufacturer's instructions. A VICTOR Nivo Multilabel plate reader (PerkinElmer, Waltham, MA, USA) was used to read the absorbance signal, setting a primary wavelength of 450 nm for all the kits. The results were converted to numeric values using standard curves.

Cell supernatant was collected, and cytokine production was analyzed with Human TNF-α ELISA kit (Invitrogen, cat. no. KHC3011), Human IL-1β ELISA kit (Invitrogen, cat. no. BMS224–2), Human IL-4 ELISA kit (Invitrogen, cat. no. BMS225–2), Human IL-6 ELISA kit (Invitrogen, cat. no. BMS213–2), Human IL-8 ELISA kit (Invitrogen, cat. no. BMS204–3), Human IL-10 ELISA kit (Sigma-Aldrich, cat. no. RAB0244), and Human IL-12p70 (Invitrogen, cat. no. KHC1578), following the manufacturer's instructions. A VICTOR Nivo Multilabel plate reader was used to read the absorbance signal, setting a primary wavelength of 450 nm for all the kits. The results were converted to numeric values using standard curves.