Il 23 quantikine elisa kit

Manufactured by R&D Systems

Sourced in United States

The IL-23 Quantikine ELISA kit is a quantitative sandwich enzyme immunoassay designed for the measurement of human interleukin-23 levels in cell culture supernates, serum, and plasma. The kit utilizes a microplate pre-coated with a monoclonal antibody specific for IL-23.

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3 protocols using il 23 quantikine elisa kit

IL-23 Quantification in THP-1 Cells

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The amount of IL-23 secreted from THP-1 cells was measured using a commercially available IL-23 Quantikine ELISA kit (R&D Systems, Minneapolis, MN, USA) following the manufacturer’s instructions. Recombinant standards provided in the kit and isolated culture media were added to wells of the plate provided in the kit. After incubation for 2 h, wells were washed and incubated with the conjugate at room temperature for 2 h. After several washes, the substrate solution was added, and the color intensity was measured. A standard curve was used for determination of the amount of IL-23 present in the samples. Data are expressed as average ± standard deviation of triplicate experiments.

Seo H.C., Kim S.M., Eo S.K., Rhim B.Y, & Kim K. (2015). 7α-Hydroxycholesterol Elicits TLR6-Mediated Expression of IL-23 in Monocytic Cells. Biomolecules & Therapeutics, 23(1), 84-89.

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Molecular Profiling of IL-23 Signaling

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Primary antibodies, such as rabbit anti-LC3B; rabbit anti-AKT, rabbit anti-p-AKT (Ser473), rabbit anti-mTOR, rabbit anti-p-mTOR (Ser2448), rabbit anti-p70S6K, rabbit anti-p-p70S6K (Thr389), mouse anti-S6, rabbit anti-p-S6 (Ser235/236), rabbit anti-NF-κB p65, rabbit anti-p-NF-κB p65 (Ser536), rabbit anti-Stat3, rabbit anti-p-Stat3 (Ser727), and rabbit anti-β-actin were obtained from Cell Signaling Technology (USA). HRP-labeled goat anti-rabbit IgG, HRP-labeled goat anti-mouse IgG, PE-labeled goat anti-rabbit IgG secondary antibodies, anti-IL-23, and anti-IL-23R were from Santa Cruz (USA). IFN-γ and TNF-α were from PeproTech (USA). Recombinant human IL-23 protein and human IL-23p19 antibody, as well as the IL-23 Quantikine ELISA kit were from R&D systems (USA). The ROS assay kit was from Beyotime Biotech (China). Rapamycin and LPS were purchased from Sigma-Aldrich (USA). IL-23R was from Abcam. Stattic was from Merck Millipore (USA), and the IKK-16 was from Selleck Chemicals (USA).

Zheng T., Xu C., Mao C., Mou X., Wu F., Wang X., Bu L., Zhou Y., Luo X., Lu Q., Liu H., Yuan G., Wang S., Chen D, & Xiao Y. (2018). Increased Interleukin-23 in Hashimoto’s Thyroiditis Disease Induces Autophagy Suppression and Reactive Oxygen Species Accumulation. Frontiers in Immunology, 9, 96.

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Quantification of Supernatant IL-23

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Supernatant IL-23 levels were measured by a quantitative enzyme immunoassay technique with the IL-23 Quantikine ELISA Kit according to the manufacture’s instructions (R&D System) after treatments with IFN-γ, LPS, or TNF-α for 24 h. A microplate reader (Thermo Fisher Scientific) capable of measuring absorbances at 450 and 570 nm was used to measure the color intensity that developed in each well. All assays were done in duplicate. The detection limit of the assay was 2.7 pg/mL.

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