SEP 22, 2016 04:00 AM PDT

Cross-validation of Antibodies Using DigiWest

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  • Head Assay Development, NMI Natural and Medical Sciences Institute at the University of Tuebingen
      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 Project Manager, Cell Signaling Technology
        Dr. Couvillon has worked at Cell Signaling Technology, Inc. for almost ten years. He joined the company as a product development scientist where he oversaw the validation and release of over 100 different antibody products. Dr. Couvillon also lead a team responsible for the generation of antibodies and kits used for enrichment and analysis of post-translational modifications. He currently works as a special project manager in the marketing group where he is committed to guiding CST's adherence to antibody validation standards and practices. He has been active in attending and presenting at major meetings regarding the topic of antibody reproducibility and works with subject matter experts, antibody users and CST's scientists to ensure that CST continues to lead the field in antibody quality and consistency.

      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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