Tansey Stool Meso Scale Discovery

Protein was isolated from human stool from individual living with Parkinson's disease (PD), inflammatory bowel disease (IBD), or neurologically healthy controls (NHC) samples using MSD lysis buffer (MSD; R60TX-3), 1 tablet complete protease inhibitor (Roche) and 5 mm stainless steel bead (Qiagen). After solids were removed by separating protein supernatant, protein concentration was determined via BCA Protein Assay Kit (Pierce), according to manufacturer instruction. Duplicates of stool (25ul) were diluted 1:1 and used to quantify chemokine and cytokines via U-PLEX custom pro-inflammatory human panel (Eotaxin, Eotaxin-3, IFN-γ, IL-1β, IL-6, IL-8, CCL22 (MDC), TNF-α (MSD#; K15067M-1) and R-PLEX human ferritin (MSD#; F21ADA-3) on the Quickplex MSD instrument, according to the manufacturer’s protocol.