Mouse cytokine chemokine luminex 25plex assay

The cytokine and chemokine panels were measured in 25 μL pancreatic lysates of 21 week old mice in duplicates using the mouse Cytokine/Chemokine Luminex 25plex Assay with a detection limit of 3.2 pg/mL to 10,000 pg/mL (EMD Millipore), as described previously (38 (link)). TGF-β levels were determined using 50 μL of pancreatic lysates in duplicates by a U-plex TGFβ Combo mouse kit with a detection limit of 37–56,600 pg/mL (TGF- β1), 2.5–45,400 pg/mL (TGF- β2), and 2.5–45,800 pg/mL (TGF- β3) (Meso Scale Discovery). For statistical evaluation, lowest detection limit was used for samples that had values below the detection sensitivity. Cytokine levels were normalized to total protein concentration measured by BCA Assay (Pierce).