Il 8 elisa kit

To measure the production of CXCL8 in tissues, we collected samples from pig lungs. These samples were then ground and processed to create a tissue suspension for detection. To measure the production of CXCL8 by PAMs, cells were transfected with lentiviral-CAST/siRNA for 48 h. Subsequently, cell supernatants were collected. The quantification of CXCL8 was performed using an IL-8 ELISA Kit (Invitrogen, USA), in strict accordance with the manufacturer’s instructions.

Conditioned medium from the cell cultures was collected at the end of each incubation, centrifuged at 14.000 × RPM for 20 min and stored at −80 °C until use. IL-8 ELISA Kit (Invitrogen, Waltham, MA, USA) was used to measure the concentration of IL-8 released in the medium, according to the manufacturer’s instructions.

To evaluate interleukin-8 (IL-8) release and cytotoxicity after incubation of cells with cathelicidin LL-37, ceragenin CSA-13 and synthesized nanoparticles MNP@LL-37 and MNP@CSA-13, cells were seeded at a density of 5 × 10⁴ cells per well in 1 mL of growth medium in 24 well plates. After 24 h of culture, the cells were treated with the tested agents at a concentration range of 1–250 μg/mL and IL-8 and LDH assays were performed. IL-8 concentration was determined using an IL-8 ELISA kit (Invitrogen). The experimental averages are shown as percent cell viability compared to control, while cells treated with lytic solution were considered as 100% LDH release.

The Multi-Analyte Profiler ELISArray Kit (SABiosciences, Frederick, MD, USA) and IL-8 ELISA Kit (Invitrogen, CA, USA) were used to determine levels of IL-1α, IL-6, IL-8, and TNF-α, according to the manufacturer's protocols. Absorbance was determined using a LabSystems Multiskan MS Analyser (Thermo Bio-Analysis Japan, Tokyo, Japan). The results were confirmed in three independent experiments.

ELISA analysis was conducted as previously described [26 (link)]. Specifically, an IL-8 ELISA Kit (Invitrogen, Carlsbad, CA, USA) was used to measure the levels of IL-8 according to the manufacturer’s protocol. The absorbance was measured using a Labsystems Multiskan MS Analyzer (Thermo Bio-Analysis Japan, Tokyo, Japan). The results were confirmed by repeating the same experiment three times.

Cells were cultured in the presence or absence of ATP, and the supernatant was collected after different treatment. The supernatant was stored at −80°C after centrifuging at 12000 rpm for 15 min at 4°C. The IL-8 and MMP-3 protein levels were measured separately using the IL-8 ELISA kit (Invitrogen, Carlsbad, CA, USA) and MMP-3 ELISA kit (Boster, Wuhan, China) following the manufacturer’s instructions. The concentration of IL-8 and MMP-3 were determined by comparing absorbance with the standard protein, and then normalized to total protein.

The IL-8 concentration in the basal media was assessed using an IL-8 ELISA kit (Thermo Fisher Scientific, Cat #88-8086-22) following the manufacturer’s protocol. The basal medium was diluted at 1:5 (without LPS/TNF-α) or 1:50 (with LPS/TNF-α) to obtain the measured signals within the dynamic range of the kit.