Dilute peptides, proteins, protein complexes, or other antigens in PBS-0.1% Tween20 to 0.1 - 0.2 mg/mL. Transfer diluted antigens transferred to 384-well print plates (15 μL/well). Antibody or other proteins/peptides prelabeled with Cy-3 and / or Cy-5 (diluted to 0.1 μg/μL) can be added to the print plate to serve as marker spots to orient the arrays post-scanning. Using a biorobotic arrayer (MicroGrid 610) and quill pins (Genomic Solution) to print antigens on nitrocellulose coated microarray slides (FAST Slide, 16 pad, Whatman Inc). Slides were kept at 75% humidity chamber for 12 hours. Store arrays at 4°in slide boxes sealed in plastic heat-sealable pouches. Reactivity is generally best within 1 month of printing, but printed antigens can retain reactivity for months.
