Zahr cytokines in pg-per-ml

These are the raw, pg/mL data from the shortly to be published PlosOne paper: Peripheral TNFα elevations in Alcohol Use Disorders are associated with Hepatitis C Infection, Natalie M. Zahr, Priya Asok, Edith V. Sullivan, & Adolf Pfefferbaum. Based on the recommendations of the Human Immune Monitoring Center (HIMC), http://iti.stanford.edu/himc.html, we used mean florescence intensity (MFI) in the manuscript. However, here we release the pg/mL data as that seems to be more useful for general use. Whole blood samples (n=223), collected in lavender EDTA tubes between March 2013 and October 2016 were centrifuged (500 rcf at room temperature for 10min). Plasma was transferred to 1.5mL conical tubes, centrifuged at 13,000 rcf at room temperature for another 10min, and the resulting supernatant was transferred to 1.5mL conical tubes for storage at −80° C until analysis by the HIMC. The HIMC, which continually benchmarks processes to minimize technical variability (Maecker et al., 2005), performed immunological assays. Human 41-plex kits (HCYTOMAG-60K, 7 kits, each able to run 42 samples) were purchased from EMD Millipore and used according to the manufacturer’s recommendations with modifications as described. Briefly, samples were mixed with antibody-linked magnetic beads on a 96-well plate and incubated overnight incubation at 4°C with shaking. Cold and Room temperature incubation steps were performed on an orbital shaker at 500-600 rpm. Plates were washed twice with wash buffer in a Biotek ELx405 washer. Following one hour incubation at room temperature with biotinylated detection antibody, streptavidin fluorochrome (i.e., streptavidin-PE) was added for 30 minutes with shaking. Plates were washed as above and PBS added to wells for reading in the Luminex 200 Instrument with a lower bound of 50-100 beads per sample per cytokine. Each sample was measured in duplicate. Custom assay control beads by Radix Biosolutions were added to all wells.