Multiplexed Nucleic Acid Programmable Protein Arrays

Yu, Xiaobo; Song, Lusheng; Petritis, Brianne; Bian, Xiaofang; Wang, Haoyu; Viloria, Jennifer; Park, Jin; Bui, Hoang; Li, Han; Wang, Jie; Liu, Lei; Yang, Liuhui; Duan, Hu; McMurray, David N.; Achkar, Jacqueline M.; Magee, Mitch; Qiu, Ji; LaBaer, Joshua

After proteins were expressed on M-NAPPA, the arrays were blocked with 5% milk-PBST for 1 h and then incubated with sera at 1:300 dilution in 5% milk-PBST for 16 h at 4 °C. After washing three times with PBST, the resulting arrays were incubated with Alex Fluor 555 labeled anti-human IgG antibody (Jackson ImmunoResearch Laboratories, PA) 1 h at 23 °C. The slides were washed with PBST, briefly rinsed with water, and dried by centrifugation (2,000 rpm, 2 min). The fluorescent scanning was performed using a Tecan scanner (Männedorf, Switzerland). The antibody binding event was quantified by fluorescence signal intensity using Array-Pro Analyzer (Media Cybernetics) software as previously reported [20, 33] .