Human il 10 uncoated elisa kit

The levels of IL‐6, IL‐10, and TNF‐α in the supernatants were determined using the following　commercial enzyme‐linked immunosorbent assay (ELISA) kits according to the manufacturer's protocol; IL‐6 Human Uncoated ELISA Kit (88‐7066‐88), IL‐10 Human Uncoated ELISA Kit (88‐7106‐88), and TNF alpha Human Uncoated ELISA Kit (88‐7346‐88) (all from Invitrogen).

Serum TGF-β1, IL-10, IL-6sR were measured with the Human/Mouse TGF-beta1 Uncoated ELISA Kit (Invitrogen, Vienna, Austria), Human IL-10 Uncoated ELISA Kit (Invitrogen), DuoSet Human IL-6Rα (R&D Systems, Minneapolis, MN, USA), respectively, according to the manufacturers’ instructions. The serum dilution factor 1/6 was chosen after titration. The readings were taken in triplicate, and the optical densities (OD) read at 450 nm with a Synergy™ H1 Microplate Reader (BioTek, Winooski, VT, USA) were compared with the standard curves prepared using recombinant proteins.

The human IL-10 uncoated ELISA kit including IL-10 protein, IL-10 capture antibody, and IL-10 detection antibody was purchased from Invitrogen. Gold (III) chloride hydrate (HAuCl₄), sodium borohydride (NaBH₄), L-ascorbic acid, 11-mercaptoundecanoic acid (MUA), and bovine serum albumin (BSA) were purchased from Sigma-Aldrich. 1-Ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride (EDC) and N-hydroxysulfosuccinimide (NHSS) were purchased from Thermo Fisher Scientific Inc. Hexadecyltrimethylammonium bromide (CTAB) was purchased from Tokyo Chemical Industry Co., Ltd. Phosphate-buffered saline (PBS) for biofunctionalization of AuNC was purchased from Biosesang.

Supernatants of M2MQs were assayed for cytokines and other released proteins. An electrochemiluminescence multiplex assay (U-Plex, Biomarker Group 1 Assays, Meso Scale Discovery (MSD), Rockville, MD, USA) was used to simultaneously quantify M-CSF, IL-1RA, TARC, IL-6, MCP-1, Eotoxin-2, IL-23, MDC, IL-13 and IL-4. The protocol and data quantification using an MSD 1300 reader was run following the manufacturer’s instructions. Enzyme-linked immunosorbent assays (ELISAs) were used for quantitation of IL-10 using Human IL-10 uncoated ELISA kit and TGFβ1 using Human/Mouse TGFβ1 uncoated ELISA kit (both Invitrogen) in Maxisorp NUNC-Immuno flat bottom 96-well plates (Thermofisher).
Fibroblast supernatants were assayed for Pro-collagen I using DuoSet ELISA Human Pro-Collagen I a1/COLIA1 kit, fibronectin using DuoSet Human Fibronectin and IL-6 using DuoSet Human IL-6 ELISA kit with the DuoSet Ancillary Reagent Kit 2 (all from R&D Systems).