Ip 10

Manufactured by Bio-Rad

Sourced in United States

IP-10 is a lab equipment product designed for the detection and quantification of the cytokine IP-10 (Interferon-Gamma Induced Protein 10). The product utilizes an enzyme-linked immunosorbent assay (ELISA) technology to provide accurate and reliable measurements of IP-10 levels in biological samples.

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5 protocols using ip 10

Serum Cytokine Detection Protocol

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For serum cytokine detection, blood samples were collected together with CRP and PCT samples at time points 0, 12, 24, 48, 72, and 96 h after study inclusion. The serum samples were kept at −75 °C until they were analyzed at the Virus Diagnostics Laboratory, University of Turku. Serum cytokine levels were determined by 27-plex immunoassay (IL-1β, IL-1rα, IL-2, IL-4, IL-5, IL-6, IL-7, IL-8, IL-9, IL-10, IL-12 (p70), IL-13, IL-15, IL-17, eotaxin, FGF basic, G-CSF, GM-CSF, IFN-γ, IP-10, MCP-1, MIP-1α, platelet-derived growth factor (PDGF-ββ), MIP-1, RANTES, TNF-α, vascular endothelial growth factor (VEGF)) from Bio-Rad Laboratories, Inc. (California, United States)). According to the manufacturer’s instructions, except that the amount of beads, detection antibodies, and streptavidin-phycoerythrin conjugate were used at 50% of their recommended concentration, which was tested previously as appropriate for the analysis system. The results were analyzed with Bio-Plex Manager 6.0 software. For statistical analyses and calculating the geometric mean cytokine levels, samples under the detection limit were given a value that was the detection limit divided by two. This was done in order to enable the inclusion of negative values (0 values) for geometric mean calculations.

Karhu J., Ala-Kokko T.I., Vuorinen T., Ohtonen P., Julkunen I, & Syrjälä H.T. (2018). Interleukin-5, interleukin-6, interferon induced protein-10, procalcitonin and C-reactive protein among mechanically ventilated severe community-acquired viral and bacterial pneumonia patients. Cytokine, 113, 272-276.

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Cytokine Profiling of LPS-Stimulated Monocytes

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LPS isolated from Escherichia coli (0111:B4) was purchased from Sigma–Aldrich (St. Louis, MO, USA). Multiplex beads for: IL-1β, IL-2, IL-4, IL-6, IL-8, IL-10, IL-12 (p70), IL-17, G-CSF, GM-CSF, IFN-γ, MCP-1, MIP-1α/1β, RANTES, TNF-α, VEGF, IP-10, and GRO-α were purchased from Bio-Rad Laboratories (Hercules, CA, USA). TGFβ was purchased from Peprotech (Rocky Hill, NJ, USA) and neutralizing TGFβ Ab was purchased from R&D (Minneapolis, MN, USA). For flow cytometry PE-conjugated anti human-CD14 and FITC-conjugated anti human-CD16 Abs purchased from eBioscience (San Diego, CA, USA) were used at a 1:50 dilution.

Aldo P.B., Racicot K., Craviero V., Guller S., Romero R, & Mor G. (2014). Trophoblast induces monocyte differentiation into CD14+/CD16+ macrophages. American journal of reproductive immunology (New York, N.Y. : 1989), 72(3), 270-284.

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Cytokine Profile of IL-17 and TNF-α Treated HDMECs

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HDMECs were starved in EBM supplemented with 2% fetal bovine serum for 6 hours and then untreated or treated with 10 ng/ml IL-17 or TNF-α or both in EBM plus 2% fetal bovine serum for 24 hours. Conditioned medium was collected and analyzed by means of xMAP technology using a X200 Luminex platform (Bio-Plex) equipped with a magnetic workstation. Panel used was the PRO Human Cytokine 27-PLEX for the simultaneous detection of FGF basic, Eotaxin, G-CSF, GM-CSF, IFN-γ, IL-1β, IL-1ra, IL-2, IL-4, IL-5, IL-6, IL-7, IL-8, IL-9, IL-10, IL-12 (p70), IL-13, IL-15, IL-17A, IP-10, MCP-1 (MCAF), MIP-1α, MIP-1β, PDGF-BB, RANTES, TNF-α, and VEGF-A (BioRad) according to the manufacturer's instruction. Data were analyzed using Bio-Plex Software Manager 6.1. Duplicate wells were used for each condition. Coefficient of variation (CV) of measurements of the whole panel was always lower than 10%.

Mercurio L., Failla C.M., Capriotti L., Scarponi C., Facchiano F., Morelli M., Rossi S., Pagnanelli G., Albanesi C., Cavani A, & Madonna S. (2020). Interleukin (IL)-17/IL-36 axis participates to the crosstalk between endothelial cells and keratinocytes during inflammatory skin responses. PLoS ONE, 15(4), e0222969.

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Investigating HO-1 Inhibitor Pyrogallol and ML385 Effects

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Pyrogallol (purity > 99.98%; HY‐N1579) (Figure 1A) and ML385 (HY‐100523) were obtained from MedChemExpress. The HO‐1 inhibitor ZnPP was purchased from AdooQ BioScience (Nanjing, China). MTT was purchased from Sigma‐Aldrich. DAPI (C1006) was obtained from Beyotime Biotechnology, Inc. (Shanghai, China). Recombinant human IFN‐β (#300‐02BC) was produced by Peprotech, Inc. (Rocky Hill, NJ, USA). Bead‐based multianalyte profiling kits for quantification of MIP‐1α, TNF‐α, MCP‐1, IL‐8, IP‐10, TRAIL, IL‐6, and RANTES were procured from Bio‐Rad Laboratories Inc. (Hercules, CA, USA). Antibodies are shown in Table S1.

Zhou B., Wang L., Yang S., Liang Y., Zhang Y., Liu X., Pan X, & Li J. (2024). Pyrogallol protects against influenza A virus‐triggered lethal lung injury by activating the Nrf2–PPAR‐γ–HO‐1 signaling axis. MedComm, 5(4), e531.

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Anticancer Peptide Effects on Gastric Cancer

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The anticancer active peptide used in the present study, anticancer bioactive peptide (ACBP) is a low-molecular-weight active substance that was obtained from the Clinical Medical Research Center, Affiliated Hospital of Inner Mongolia Medical University (Hohhot, China). The poorly differentiated human gastric adenocarcinoma cell line of MKN-45, the highly differentiated human gastric adenocarcinoma cell line NCI-N87 and the immortalized and non-tumorigenic human gastric mucosal epithelial cell line GES-1 were purchased from the Cell Bank of the Chinese Academy of Sciences (Shanghai, China). RPMI-1640 medium and high-glucose DMEM were purchased from Hyclone (Cytiva). Penicillin, streptomycin, fetal bovine serum and 0.25% trypsin were from Gibco (Thermo Fisher Scientific, Inc.).
The Bio-Plex ProT Human Cytokine 27-plex Assay (cat. no. M500KCAFOY; Bio-Rad Laboratories, Inc.) is a multiplex assay that detects cytokines including IL-1β, IL-1RA, IL-2, IL-4, IL-6, IL-7, IL-8, IL-9, IL-10, IL-12, IL-13, IL-15, IL-17, Eotaxin, bFGF, GM-CSF, IFN-γ, MCP-1 (MCAF), IP-10, MIP-1α, PDGF-BB, MIP-1β, RANTES, TNF-α and VEGF. OXA was obtained from Jiangsu Hengrui Pharmaceutical Co., Ltd.

Li X., Peng J, & Su X. (2023). Expression of immune regulatory factors, chemokines and growth factors in differentiated gastric cancer cells treated with an anticancer bioactive peptide combined with oxaliplatin. Molecular and Clinical Oncology, 20(1), 9.

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