Human tgfβ1 elisa kit

To quantify plasma cytokine levels, the multiplex ELISA-based Q-Plex™ Human Cytokine HS Screen Array (Quansys Biosciences, Logan, Utah, USA) was used. The kit contains an array of inflammatory cytokines, including IL-1α, IL-1β, IL-2, IL-4, IL-5, IL-6, IL-10, IL-12p70, IL-13, IL-15, IL-17, IL-23, IFN-γ, TNF-α, and TNF-β. The assay was performed in a 96-well plate following the manufacturer’s protocol. The Q-View™ Imager Pro and Software were used for data capture and calculation of the final concentrations, respectively. Additionally, human TGF-β1 ELISA Kit (BOSTER Biological Technology, Pleasanton, CA, USA) was used to quantify plasma TGF-β levels. Briefly, the amount of TGF-β was determined at an optical density of 450 nm wavelength through the 800 TS Absorbance Reader (Bio-Tek instruments Inc., Winooski, VT, USA). The TGF-β concentrations of MPN samples were calculated through interpolation against a standard curve. In all samples, the plasma levels of various cytokines were analyzed in duplicates. Samples with low precision and repeatability (defined as a coefficient of variation of more than 10%) and those with apparent outlier values were excluded from subsequent statistics.

A human complement protein C3 ELISA kit (#CSB-E08665h, Wuhan Huamei, China) was used to measure C3 according to the manufacturer’s instructions. A human TGFβ1 ELISA kit was used to measure TGFβ1 in plasma using a previously described procedure [14 (link)]. A human TGFβ1 ELISA kit (#EK0513, Boster, China) was used to measure TGFβ1 in urine following the manufacturer’s instructions.

IFN-γ and TGF-β1 were measured by ELISA assay on the culture supernatant collected on day 5 from suppression experiments. In particular, cytokine concentration was assessed by Human IFN-gamma Instant ELISA (Bender MedSystems) and Human TGF-β1 ELISA kit (Boster Biological Technology Co). Samples were acquired by LB 940 Multimode Reader Mithras (Berthold Technologies).