Antibody microarray analysis using twocolor rolling circle amplification

All assays were performed as described in Zhou et al. [10]. Briefly, the labeled samples (different serum types) were incubated on antibody microarrays containing 48 unique antibodies that were printed on NC-coated glass microscope slides (FASTTM slides; Schleicher & Schuell Bioscience, Keene, NH USA). Labeling was performed by mixing the serum samples 1:15 with 50 mM carbonate buffer, pH 8.3, and 1/20th volume 6.7 mM N-hydroxysuccinimide (NHS) ester linked biotin or digoxigenin (Molecular Probes-Invitrogen) in DMSO. The reactions were quenched and unreacted dye was removed by size exclusion spin chromatography (Bio-Spin-P6, Bio-Rad), molecular weight cutoff = 6000 Da). The arrays were detected using the TC-RCA procedure, as detailed in [10].

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