Legendplex rat th cytokine panel 13 plex

At post-mortem, blood was collected from the heart and placed in Serum Gel-Z tubes (Sarstedt, Numbrecht, Germany), which were then centrifuged at 2000 rpm for 5 min. Serum was collected and stored at −80 °C until used for cytokine analysis. Quantitative analysis of cytokines was performed using a LEGENDplex Rat Th Cytokine Panel (13-plex; Biolegend, San Diego, CA, USA), following the manufacturer’s instructions. Data were acquired using a FACSCanto II flow cytometer (BD Biosciences, San Jose, CA, USA) and analysed using Legendplex software (Biolegend).

We analyzed plasma samples using a LEGENDplex Rat Th Cytokine Panel (13-plex) (Biolegend, San Diego, CA), which allowed simultaneous analysis of 13 cytokines: Interleukin (IL)-2, IL-4, IL-5, IL-6, IL-9, IL-10, IL-13, IL-17A, IL-17F, IL-22, granulocyte-macrophage colony-stimulating factor (GM-CSF), interferon (IFN)-γ and tumor necrosis factor (TNF)-α. The cytokine kit was modified for use with 384-well plates and the samples processed, as described previously [30 (link)]. Many of our IL-6 samples showed levels far above the upper limits of our established flow-cytometry methods. To compensate, we re-ran the saturated samples using a further dilution factor of 50:1.