Antibody arrays are used to measure different proteins simultaneously in one experiment, with significantly less time, effort and cost than running the many enzyme-linked immunosorbent assays (ELISAs) or western blots (WBs) required for the same number of analytes. Creative Biolabs has established an advanced antibody array technology platform for customized antibody array development for in vitro diagnostics (IVD) research use.
Antibody arrays, also known as antibody microarray, allows for the screening or profiling of multiple proteins within a sample entireproteome. Antibody arrays are an antibody-pair-based assay, analogous to ELISA, but using the membrane or glass as a substrate rather than a plate. Proteins are extracted from samples and labeled with the appropriate conjugate. After cleanup to remove any unbound, free label, the proteins are allowed to bind to antibodies captured on the array. Once bound, the array can be visualized for those proteins labeled with a biotin-fluorophore. If proteins were labeled with biotin alone, visualization is done through fluorescence or chemiluminescence, usually involving a streptavidin conjugate. Each array includes positive, negative and internal controls. Redundancy in the number of antibodies spotted onto the array ensures effective and accurate results.
Rolling-circle amplification (RCA) can greatly enhance fluorescence signals over non-amplified detection, enabling the detection of low-abundance proteins like cytokines. A schematic representation of a two-color version of the method is shown in Figure 1.
Fig.1 Schematic representation of two-color RCA on antibody microarrays. (Haab, 2006)
The biological roles of proteins are determined not just by their abundance, but also by modifications and interactions that occur after translation such as phosphorylation, glycosylation, cleavage, oxidation or reduction, and lipid attachment. Schematic representations of 'heterogeneous' sandwich assays are shown in Figure 2 for the measurement of glycosylation and phosphorylation states.
Fig.2 The detection of phosphorylation and glycosylation on antibody arrays. (Haab, 2006)
If you are interested in our custom antibody array development services, please contact us for more information.
Reference
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