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.
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Antigen Arrays: Technical Details
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.
