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An Antibody Based Proteomics Discovery Platform
Post-translational modifications (PTMs), such as phosphorylation, acetylation, methylation, or ubiquitination, are critical regulators of protein activity and function. Understanding the role of PTMs in disease states like cancer and developing therapeutics to combat their decoupled regulation are areas of intense research. Cell Signaling Technology (CST) has developed a proprietary proteomic method, PTMScan® Technology, that overcomes this challenge by using immunoaffinity enrichment to isolate specific modifications of interest from a protease-digested cell or tissue extract.
Following a general introduction to the technology and a review of the PTMScan protocol the attendees will perform the method with guidance from CST scientists including peptide preparation, immunoaffinity enrichment, and C18 tip sample cleanup to prepare samples for LC-MS/MS analysis. Cell Signaling Technology (CST) will provide all reagents to perform these steps, including the option for attendees to choose from Phosphotyrosine, Ubiquitin K-GG Remnant, or Acetyl-Lysine Motif Antibodies for their enrichments. Once prepared, samples can be taken back to attendee labs for LC-MS/MS analysis and comparison to CST-generated data that will be provided for each antibody.
In addition to the practical biochemistry component, the workshop will cover LC-MS/MS instrument settings, approaches to data analysis, and common software tools used to analyze the LC-MS/MS data.
The workshop will also include a number of examples from the published literature of applications of PTMScan. Here we will review hallmark discoveries published using PTMScan, including the use of anti-phosphotyrosine antibodies to discover drivers of malignancy in cancer; how ubiquitin K-GG remnant antibodies were used to uncover mechanism of action of viral infection, or how protein stability can regulate cellular development.
Charles L. Farnsworth, Ph.D.
Senior Proteomics Application Scientist
Brief summary of cv:
B.S. biology, Boston University, Boston, MA
Ph.D. biochemistry, Tufts University School of Medicine, Boston, MA
Post-doctoral fellow, department Cell Biology, Harvard Medical School, Boston, MA
Fellow, Leukemia and Lymphoma Society of America, Harvard Medical School, Boston
Research scientist lll, cancer biology group, Cell Signaling Technology, Danvers, MA
Senior Application scientist, PTMScan group, Cell Signaling Technology