Antigen Array Production

• Dilute peptides, proteins, protein complexes, or other antigens in PBS or H20 to 0.1 - 0.2 mg/mL (up to 30% DMF or DMSO can be used for peptides and proteins that are insoluble in H20 or PBS).
  
  
• Transfer diluted antigens transferred to 384-well "print plates" (7 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 robotic arrayer and TeleChem International SMP7B Stealth Micro Spotting Pins (Telechem International, Sunnyvale, CA) to print antigens on Poly-L-lysine coated microarray slides (CEL Associates, Pearland, Texas). For optimal results, increased pin sonication between loads (5 x 7 second sonication cycles), between print plates (5 minutes), and at the end of each print (30 minutes) are recommended.
  
  
• 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.

Antigen Arrays: Technical Details Up to 12,000 features/array 200 mM features 50 pg Ag/feature Probe with 1:150 dilutions sera Use common array equipment Antigen Microarray site

Figure. Antigen array production. The pictured robotic arrayer, developed by, is used to attach putative peptide and protein autoantigens in ordered arrays on derivatized microscope slides.