Large-scale antibody analysis for protein phosphorylation events

Around 30% of the proteins in our antibody catalog are subject to phosphorylation, a complex post-translational modification which regulates practically all cellular processes in eukaryotes. Many human diseases can be attributed to abnormal phosphorylation events. We at have a large antibody database of phospho-antibodies, which are used to study the cell cycle, differentiation, metabolism, intercellular communication and many other cellular activities.

New answers to complex phosphorylation problems are constantly being sought, with antibody suppliers having an ever widening phosphorylation database resource to call upon as a result. Research has included sequence alignment studies to identify protein kinase genes; in vitro assays with synthetic peptides to specify kinase actions, and systematic studies to identify specific protein targets. However, research is limited due to the technical challenges of such a large-scale approach, and currently only Mass Spectrometry Proteomics has the capability to identify kinase substrates and map specific modification positions en masse.

However, a combination of high throughput cutting-edge technology; better phosphopeptide purification methods; the development of phospho-specific antibodies, and improved data-handling software has considerably improved matters. In 2007, Judit Villn et al. employed an integrated high-throughput analytical platform process to isolate and identify 5,635 phosphorylation sites, from 2,328 mouse liver proteins. From this, they were able to extract novel and known Ser/Thr kinase motifs, including a unique dipolar motif.

Antibody suppliers like us at Novus Biologicals play a large part in the advancement of phosphorylation research. As well as developing an extensive antibody database of kinase and phospho-specific antibodies, we have created a range of Mix-and-Match Pathway Assay Kits, for simultaneous detection of multiple kinases on a single 96-multiwell platform. In addition, our antibody catalog features the unique Precipitor High Throughput Immunoprecipitation and Purification system; an automated magnetic bead platform with the ability to handle 16 different assays on a 96 deep-well plate.

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