HeLa cells were seeded at a concentration of 2 X 105 cells/ml 2 days before infection. Salmonella strains were first inoculated in LB broths containing appropriate antibiotic selection and grown overnight with shaking at 37°C. Overnight cultures were then subinoculated 1/200 in fresh LB broths and grown under static conditions for additional 20 hrs at 37°C. HeLa cells were then infected at a MOI of 50 for 1 h, followed by replacing with media containing 100 μg/ml gentamycin for 1 h. HeLa cells were then further grown in the media containing 10 μg/ml gentamycin with untreated alone or the stimulation by 20 ng/ml TNFα for 6 hrs. The supernatants were collected to measure IL-8 secretion using the Quantikine Human IL-8 Immunoassay (R&D Systems, MN) according to the manufacturer’s instructions. Differences in IL-8 secretion were assessed for significance by one-way analysis of variance (ANOVA) with Turkey’s Multiple Comparison post-test.
Quantikine human il 8 immunoassay
Manufactured by R&D Systems
Sourced in France
The Quantikine Human IL-8 Immunoassay is a quantitative sandwich enzyme immunoassay designed to measure human interleukin 8 (IL-8) levels in cell culture supernates, serum, and plasma. It utilizes a microplate format and a pre-coated capture antibody specific for IL-8.
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Lab products found in correlation
3 protocols using quantikine human il 8 immunoassay
1
Salmonella Infection Induces IL-8 Secretion
2
Assessing Cytokine Production in TiO2-NP Exposed Cells
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A549 cells and co-culture cells were seeded in 96-well-plates (1x10 5 cells/well in 50 μL of medium) and were allowed to adhere for 24 h. NPs were diluted in DMEM cell culture medium to reach the following final concentrations: 15, 30, 60 and 120 μg/mL. After 24h cell exposure to TiO2-NPs, the production of Tumor Necrosis Factor alpha (TNF-α) was evaluated in coculture using a commercial ELISA Kit (Quantikine® Human TNF-α Immunoassay; R&D Systems, Lille, France) according to the manufacturer's instructions. Interleukin-8 (IL-8) production was assessed in the two cell systems (A549 cells and A549/ differentiated-THP-1 co-culture) after exposure to TiO2-NPs for 24 h by a commercially available ELISA kit (Quantikine® Human IL-8 Immunoassay; R&D Systems, Lille, France) according to the manufacturer's instructions. The optical density of each sample was determined using a microplate reader (Multiskan RC; Thermolabsystems, Helsinki, Finland) set to 450 nm. Three independent experiments were performed, the production of TNF-α and IL-8 were reported to that of control (unexposed) cells.
3
Inflammatory response to TiO2 NPs
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10 5 cells/well were seeded on 96 well plates. Cells were exposed to 15, 30, 60 and 120 μg/mL of pristine and irradiated TiO2 NPs. After 24 h exposure, the production of the pro-inflammatory markers tumor necrosis factor alpha (TNF-α), and interleukin 8 (IL-8) were determined by sandwich enzyme-linked immunosorbent assay (Quantikine Human TNF-α, and Quantikine Human IL-8 Immunoassay; R&D Systems, Lille, France) according to the manufacturer's instructions. The optical density of each sample was determined using a microplate reader (Multiskan RC; Thermo Labsystems, Helsinki, Finland) set to 450 nm. Three independent experiments were performed, and the production of TNF-α and IL-8 was reported to that of control (unexposed) cells.
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