Multiplexed protein analysis using spotted antibody microarrays.

This chapter describes methods for the production and use of antibody microarrays. The methods are divided into (a) antibody handling and microarray production, (b) sample preparation, and (c) microarray use. Two types of detection methods are described: direct labeling and a fluorescence-linked immunosorbent assay (FLISA). In the direct labeling method, all proteins in a complex mixture are labeled with either a fluorophore or a hapten that allows subsequent detection. In FLISA detection, a capture antibody on the microarray captures the unlabeled protein target, which is detected by a detection antibody and a fluorophore-labeled secondary antibody. Each method has particular optimal uses, which are discussed in the text.