Human il 6 uncoated elisa

The sera for the analysis of biomarkers were sampled on V0–V2 and stored at −80 °C until analysis. The enzyme-linked immunosorbent assays (ELISA) were performed following the sandwich ELISA principle using the ELISA reader Infinite M200 Pro, TECAN Group Ltd., Männedorf, Switzerland. The corresponding manufacturer’s instructions were followed when performing the ELISAs (Human IL-6 Uncoated ELISA; Human TREM2 ELISA Kit; Human BDNF ELISA Kit; Invitrogen, ThermoFisher Scientific, Waltham, MA, USA; Gasdermin D ELISA, Horizon Bioscience Discovery Ltd., Cambridge, UK). However, deviating from this, the standard curve of the IL-1β-ELISA (Interleukin-1beta high sensitivity ELISA; IBL International GmbH, Hamburg, Germany,) was extended by a lower concentration by adding a further dilution step (1:2). The subsequent photometric measurement of the absorbance was performed at a wavelength of 450 nm by the appropriate microplate reader (ELISA reader: Multiskan EX, Thermo Electron Corporation, Waltham, MA, USA). The generation of the standard curve and calculation of the concentrations are performed with the statistical software GraphPad PRISM 8.0.1.
Values for CRP (all measured with the Dimension Vista, Siemens Healthcare Diagnostics, Eschborn, Germany) were measured in the central laboratory facility of the University Medicine Greifswald.

To determine cytokine production, the cell supernatants were harvested 48 h post-infection, and IL-6 levels were detected using human IL-6 uncoated ELISA (Invitrogen, Carlsbad, CA, USA).

Cytokines Levels of IFN-γ, TNF-α, IL-6, and IL-10, were measured in the plasma, uninfected PBMC lysates at pre-supplementation and post supplementation in both placebo and experimental group. Cytokine levels were also measured in the mycobacteria-infected in vitro granuloma supernatants to determine the effects of in vitro PZA + in vivo GSH treatments and in vitro INH + in vivo GSH treatments in altering the levels of these cytokines. Cytokine levels were measured using enzyme-linked immunosorbent (ELISA) assay kits: Human 1L-10 ELISA Ready-SET-Go from Affymetrix (Cat# 88-7106) and Human TNF-α Uncoated ELISA (Cat # 88-7346), Human IFN-γ Uncoated ELISA (Cat # 88-7316), and Human IL-6 Uncoated ELISA (Cat # 88-7066) from Invitrogen to quantity detection of cytokines. Assays were performed following the manufacturer’s protocol for each kit.