SEP 22, 2016 04:00 AM PDT
Cross-validation of Antibodies Using DigiWest
3 19 1960

Speakers:
  • Head Assay Development, NMI Natural and Medical Sciences Institute at the University of Tuebingen
    Biography
      Dr. Templin heads the Department of Assay Development at the NMI - Natural and Medical sciences Institute at the University of Tübingen. In a multidisciplinary environment, scientists from applied physics, chemistry, biology and biochemistry are working together in his department to develop advanced assay system for industrial clients.
      Markus Templin's research is focused on the application of microarrays for proteomic and cancer research and he holds several patents in this area. He studied biology at the University of Tübingen, did his postdoctoral research at Edinburgh University, Scotland and the Max-Planck-Institute for Developmental Biology, Germany before joining the Natural and Medical Sciences Institute.
    • Scientific Marketing Project Manager, Cell Signaling Technology
      Biography
        Dr. Couvillon attended Colby College in Waterville, Maine (USA) where he majored in Biology and Chemistry. Post-graduation, he became a research associate in the laboratories of Dr. Christopher L. Carpenter and Lewis Cantley at the Harvard Medical School where he was introduced to the fascinating world of signal transduction, lipid kinases and the role of Ras-like proteins in disease. Under the tutelage of Dr. John H. Exton at Vanderbilt University (Nashville, Tennessee), Dr. Couvillon received his Ph.D. in Molecular Physiology and Biophysics while studying the role of Rho-family proteins and their regulators in neuronal function. He then returned to Harvard and Beth-Israel Deaconess Medical Center where he rejoined the Cantley lab to continue his work on Rho-family GTPases and their role in immune cell development and function, neuronal plasticity and cardiac development. Dr. Couvillon joined Cell Signaling Technology in 2009 as a Development Scientist, developing and validating antibody products in the MAPK, lipid signaling and other pathways. He has also worked extensively in the proteomics space, developing novel antibody reagents against novel post-translational modification and reagents that detect specific substrate motifs for high-throughput proteomics analysis. Dr. Couvillon currently works as a Scientific Project Manager focused on initiatives promoting antibody reproducibility in research.

      Abstract:
      DATE: September 22, 2016
      TIME:  4:00am PT, 7:00am ET, 11:00am GMT


      Here, we present a novel approach that combines the principles of the western blot (protein separation by SDS-PAGE; protein immobilization on a solid support; detection by specific antibodies with a multiplexed bead array as a readout system. The system is able to generate of hundreds of bead-based western blot equivalents from a few micrograms of protein, allowing the user to obtain information on the expression and modification of hundreds of proteins from a small sample. This new approach is highly reproducible and has both good linearity and a large dynamic range.
      Primary antibodies are the most critical component of both conventional western blots and DigiWest. Therefore, good antibody validation dedicated to each application is key to good and reliable results. During this webinar we will show what a good antibody validation protocol means and how this differs in different applications.

      Taking advantage of the capabilities of DigiWest, we performed comprehensive signal transduction analysis of a lung cancer cell line that was rendered resistant to the kinase inhibitor lapatinib. To analyze the activation state of different signaling cascades hundreds of antibodies were employed while only micrograms of protein were required. Major changes in the activation state of Ephrin-mediated signaling and a central role for p53-controlled processes that form the basis of the observed resistance could be revealed. 
       

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