Immunostaining with dissociable antibody microarrays

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Abstract
The availability of a large number of biological materials such as cDNA, antibodies, recombinant proteins, and tissues has promoted the development of microarray technologies that make use of these materials in high‐throughput screening assays. However, because microarray technologies have been less successful in examining proteins than DNA and mRNA , there is a need for improved protein microarray systems. To address this need, we developed an antibody microarray‐based immunostaining method that can analyze the properties of a large number of proteins simultaneously. In this method, antibodies are arrayed and immobilized on a solid support and cells bearing antigens of interest are attached to a second support. Apposition of the two supports allows the antibodies to dissociate from the array support and bind to the cellular antigens. After separation of the supports, antigen‐bound antibodies can be detected by standard secondary antibody techniques. These “dissociable” antibody arrays were used to detect both the expression and subcellular localization of a large number of specific proteins in various cultured cell types.