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Peptide Microarrays for Profiling of Modification State-Specific Antibodies

Zerweck et. al., Methods in Molecular Biology, Epitope Mapping Protocols (2009) - PMID: 19377944

Product(s) used in this publication:  PepStar™ Peptide Microarrays

Abstract:

The reversible phosphorylation of serine, threonine, and tyrosine residues is one of the most important intracellular post-translational modifications regulating enzymatic activities and protein/protein interaction in eukaryotic cells. Tools for determining phosphorylation status of proteins and peptides play a prominent role in signal transduction research and proteomics. Pan-specific antibodies claimed to recognize modified amino acid residues independent on the nature of surrounding residues in peptides and proteins are widely used. We used high-content phosphopeptide microarrays and microarrays displaying acetyllysine-containing peptides for comprehensive characterization of commercially available generic anti-phosphopeptide and anti-acetyllysine antibodies. We were able to demonstrate distinct subsite specificity and cross-reactivity for such antibodies.

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