| Description |
Serum samples were probed using HuProt v3.1 arrays (CDI Laboratories, Inc.), which comprised of more than 20,000 unique human full-length proteins, according to the manufacturer's instruction. Briefly, the HuProt array is based on the sandwich immunoassay principle. More than 20,000 unique human proteins were duplicately immobilized in specific location spots on the surface of the array membrane. After blocking, HuProt array was incubated in 3 mL 1:1,000 diluted serum sample to capture corresponding autoantibodies to each protein spot, and followed with 1:2,000 diluted Cy3-conjugated goat anti-human IgG (Jackson, 109-165-008) and Alexa Fluor 647-affinipure donkey anti-human IgM (Jackson, 709-605-073) to fluorescently label IgG and IgM of captured autoantibodies, respectively. Finally, the HuProt assays were scanned using the GenePix 4000B Microarray Scanner (Molecular Devices, Sunnyvale, CA, USA) and analyzed with GenePix Pro 6.0 software (Molecular Devices, Sunnyvale, CA, USA). |
