The plasma levels of IL-6 and IL-8 were determined by ELISA, following the manufacturer’s recommendations. The Human IL-6 High Sensitivity ELISA Kit from eBioscience (Waltham, MA, USA) and the Human Ultrasensitive ELISA Kit from Thermo-Fisher (Waltham, MA, USA) for IL-8 were used.
Human il 6 high sensitivity elisa kit
Manufactured by Thermo Fisher Scientific
Sourced in United States
The Human IL-6 High Sensitivity ELISA Kit is a laboratory assay used to quantitatively measure the concentration of interleukin-6 (IL-6) in human samples. It employs the quantitative sandwich enzyme immunoassay technique.
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Lab products found in correlation
Human ultrasensitive elisa kit, by Thermo Fisher Scientific (2 mentions)
Elisa kit, by R&D Systems (1 mentions)
Human tnf ri tnfrsf1a quantikine elisa kit, by R&D Systems (1 mentions)
Cytometric bead array human inflammatory cytokine kit, by BD (1 mentions)
Lsrfortessa flow cytometer, by BD (1 mentions)
Elisa kit, by Thermo Fisher Scientific (1 mentions)
5 protocols using human il 6 high sensitivity elisa kit
1
Plasma IL-6 and IL-8 Quantification by ELISA
2
Plasma IL-6 and IL-8 Quantification
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The plasma levels of IL-6 and IL-8 were determined by ELISA, following the manufacturer’s recommendations. For IL-6, we used the Human IL-6 High Sensitivity ELISA Kit from eBioscience and for IL-8, the Human Ultrasensitive ELISA Kit from Thermo-Fisher.
3
Dendritic Cell Phenotyping with ELISA
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The agreement between the two measurements (preand post-angiography) was assessed with a two-way mixed intra class correlation coefficient (ICC, absolute agreement, single measures). The ICC was interpreted based on kappa values as proposed by Landis and Koch 27 : ELISA kit, R&D systems; Human IL-6 High-Sensitivity ELISA kit, eBioscience, Vienna, Austria) according to the manufacturers' recommendations. All measurements were performed in duplicate. The sensitivity of sCD14 and IL-6 ELISA was 125 pg/ml and 0.03 pg/ml, respectively. The intra-assay coefficients of variation (CV) for both assays were set at ≤ 15%.
Fig. 2 Gating strategy for dendritic cells. CD45+ (APC-H7) cells with a SSCLowPerCPphenotype were gated as the population of interest (POI), containing CD14-CD19-live cells. This POI was gated for the presence of BDCA-1 and BDCA-2 (a). Then, a stringent gate was drawn around respectively BDCA-1 + and BDCA-2 + cells using an APC-H7/SSC density plot in order to exclude irrelevant cells (b). Figure c shows the BDCA-1 + mDC and BDCA-2 + pDC.
Fig. 2 Gating strategy for dendritic cells. CD45+ (APC-H7) cells with a SSCLowPerCPphenotype were gated as the population of interest (POI), containing CD14-CD19-live cells. This POI was gated for the presence of BDCA-1 and BDCA-2 (a). Then, a stringent gate was drawn around respectively BDCA-1 + and BDCA-2 + cells using an APC-H7/SSC density plot in order to exclude irrelevant cells (b). Figure c shows the BDCA-1 + mDC and BDCA-2 + pDC.
4
Cytokine Profiling in Inflammaging and Asthma
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Proinflammatory cytokines known to be involved in Inflammaging [4 (link)–8 ] (TNFα, IL-1β and IL-6, IL-8, IL-12 and sTNF RI) and to be important for the pathogenesis of bronchial asthma were selected for testing. The study also included IL-10, a cytokine with immunoregulatory and antagonistic properties. The serum levels of TNF-α, IL-1β and IL-6 were determined with the use of the commercial Thermo Fisher Scientific Ultrasensitive TNF alpha Human ELISA Kit, High Sensitivity IL-1 beta Human ELISA Kit and High Sensitivity IL-6 Human ELISA Kit. The serum level of soluble TNF RI was determined with the use of the commercial R&D Systems Human TNF RI/TNFRSF1A Quantikine® ELISA Kit. IL-12p70, IL-10 and IL-8 levels in the serum were measured with the BD™ Cytometric Bead Array Human Inflammatory Cytokine Kit. The samples were run on a BD™ LSR Fortessa flow cytometer and the results were analyzed with the BD™ FCAP Array software.
IL-10 was detectable only in 48/118 (40.7%), IL-12p70 in 20/118 (16.9%) and IL-1β in 10/118 (8.5%) of the study participants, and IL-10, IL-12p70 and IL-1β were not included in the analysis.
IL-10 was detectable only in 48/118 (40.7%), IL-12p70 in 20/118 (16.9%) and IL-1β in 10/118 (8.5%) of the study participants, and IL-10, IL-12p70 and IL-1β were not included in the analysis.
5
Quantification of Secreted IL-6 and IL-10
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Secreted interleukin-6 (IL-6) and IL-10 were quantified using human IL-6 and IL-10 enzyme-linked immunosorbent assay (ELISA) kits (Thermo Fisher Scientific), according to the manufacturer’s protocol. To detect IL-6 in the U2932 cell line, the high-sensitivity IL-6 human ELISA kit was used (Thermo Fisher Scientific).
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