Antibody Microarrays for Expression Analysis

In recent years, antibody microarrays have developed into an important tool for proteomics. As a multiplexing technique, they facilitate the highly parallel detection of hundreds of different analytes from very small sample volumes of only a few microlitres. This is combined with high sensitivity in the picomolar to femtomolar range, which is similar to the sensitivity of ELISA, the gold standard for protein quantification. In order to obtain such sensitivities in a robust and reproducible manner for sets of several hundreds of analytes simultaneously, it is essential to optimise the experimental layout, sample handling, labelling and incubation as well as data processing steps. Here, we present our current antibody microarray protocols for multiplexed expression profiling studies, which permit the analysis of the abundance of more than 800 proteins in plasma, urine and tissue samples.