Opteia human il 10 elisa set

Manufactured by BD

The BD OptEIA™ Human IL-10 ELISA Set is a laboratory equipment product that enables the quantitative measurement of human interleukin-10 (IL-10) levels in biological samples using the Enzyme-Linked Immunosorbent Assay (ELISA) technique.

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Close spelling variants of this product

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5 protocols using opteia human il 10 elisa set

Cytokine Quantification Using ELISA

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The collected culture media from all treatments and cell types were analyzed using an enzyme-linked immunosorbent assay for IL-1 (BD OptEIA™ Human IL-1 ELISA Set—557953, BD Biosciences, IL-6; BD OptEIA™ Human IL-6 ELISA Set—555220, BD Biosciences, DF: 1:2), and IL-10 (BD OptEIA™ Human IL-10 ELISA Set—555157, BD Biosciences), as per the kit instructions. For each ELISA, duplicate standards were used to create a dilution series according to the manufacturer’s recommendations. The assay results were analyzed using a Synergy H4 microplate reader (BIO-TEK).

Menon R., Khanipov K., Radnaa E., Ganguly E., Bento G.F., Urrabaz-Garza R., Kammala A.K., Yaklic J., Pyles R., Golovko G, & Tantengco O.A. (2023). Amplification of microbial DNA from bacterial extracellular vesicles from human placenta. Frontiers in Microbiology, 14, 1213234.

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Quantifying IL-10 Production in T Cells

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The concentrations of IL-10 in supernatant or sera were determined using BD OptEIA Human IL-10 ELISA Set (BD Biosciences). To test IL-10 production, CD8⁺ T cells were purified from PBMCs (MACS Column Technology), and labeled with tet-APC, CD8-PECy7, CD4-PE and violet. 6×10⁴ FACS-sorted cells were distributed into 96 wells with 200 μl medium containing 50 IU/ml rhIL-2, T2 cells (2:1 ratio) pulsed with peptide NY-ESO-1 157–165 or control peptide HIVpol 476-484 (10 μg/ml). Supernatant was collected for ELISA assay after 48-hour incubation. Alternatively, CD4⁺ and CD8⁺ T cells were separated by MACS Column Technology, the rest of the cells were labeled with CD14-ECD, CD11c-Alexa700, CD3-PerCPCy5.5, CD56-PE and CD19-FITC and sorted. Total RNA was extracted from each cell subset, and IL-10 mRNA was detected using RT-QPCR as previously described (43 ).

Sun Z., Fourcade J., Pagliano O., Chauvin J.M., Sander C., Kirkwood J.M, & Zarour H.M. (2015). IL-10 and PD-1 cooperate to limit the activity of tumor-specific CD8+ T cells. Cancer research, 75(8), 1635-1644.

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Quantifying IL-10 Secretion in B Cells

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B cells were stimulated as previously described. Thereafter, cell-free supernatants were harvested and IL-10 secretion was quantified using a commercial BD OptEIA Human IL-10 ELISA set (BD Biosciences).

Andreani G., Ouellet M., Menasria R., Gomez A.M., Barat C, & Tremblay M.J. (2015). Leishmania infantum Amastigotes Trigger a Subpopulation of Human B Cells with an Immunoregulatory Phenotype. PLoS Neglected Tropical Diseases, 9(2), e0003543.

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Cytokine Profiling of Immune Cells

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Culture media collected from ecto, endo, stroma, and THP-1 macrophages 48 h post-infection were tested for human GM-CSF, IL-6, IL-8, IL-10, and TNFα using the BD OptEIA^™ Human GM-CSF ELISA Set (555126, BD Biosciences, San Jose, CA), BD OptEIA^™ Human IL-6 ELISA Set (555220, BD Biosciences), BD OptEIA^™ Human IL-8 ELISA Set (555244, BD Biosciences), BD OptEIA^™ Human IL-10 ELISA Set (555157, BD Biosciences), and Human TNF ELISA Set (555212, BD Biosciences), respectively. Standard curves were developed using duplicate samples of known-quantity recombinant proteins that were provided by the manufacturer. Sample concentrations were determined by relating the absorbance of the samples to the standard curve using linear regression analysis.

Tantengco O.A., Kechichian T., Vincent K.L., Pyles R.B., Medina P.M, & Menon R. (2021). Inflammatory Response Elicited by Ureaplasma parvum colonization in human cervical epithelial, stromal, and immune cells. Reproduction (Cambridge, England), 163(1), 1-10.

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Cytokine Production from Stimulated PBMCs

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Peripheral blood mononuclear cells (1 × 10⁶ cells/mL per well) were cultured in RPMI-1640 (Gibco, Life Technologies, Foster City, CA, USA) medium added with fetal bovine serum (FBS) (Sigma, St. Louis, MO, USA) at 37°C in an atmosphere of 5% CO₂, overnight. Cells were stimulated with Staphylococcus aureus (10 to 1 microorganism per cell) after 5 hours of incubation with the test compounds. After additional 18 hours of incubation the supernatants were stored at −80°C. TNF-α and IL-10 were quantified by enzyme-linked immunosorbent assay (ELISA) using BD OptEIA Human TNF ELISA Set (Cat. no. 555212) and BD OptEIA Human IL-10 ELISA Set (Cat. no. 555157), respectively, according to the manufacturer's instructions.

Paracatu L.C., Faria C.M., Quinello C., Rennó C., Palmeira P., Zeraik M.L., da Fonseca L.M, & Ximenes V.F. (2014). Caffeic Acid Phenethyl Ester: Consequences of Its Hydrophobicity in the Oxidative Functions and Cytokine Release by Leukocytes. Evidence-based Complementary and Alternative Medicine : eCAM, 2014, 793629.

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