Workshops
Seminar rooms for workshops: N55 / SR 210/11, N55 / SR 205 & N55 / SR 206We are offering the following workshops:
Surface Plasmon Resonance-Workshop - In vitro analyses of biomolecular interactions
"Membrane- and sorting signal binding by the clathrin adaptor AP2: What we learned from structural and SPR biosensor-based functional analyses"Speaker: Stefan Höning (University of Cologne)
The topic of this rather informal seminar is the real-time analyses of biomolecular interactions with a surface plasmon resonance (SPR) biosensor. The speaker will introduce the SPR technology/principle and the technical systems available. Multiple examples of protein-protein and protein-lipid interactions will illustrate application areas and the advantages and limitations of the technology. The methods of molecule coupling/capture on a sensor surface as well as the details of kinetic measurements and data evaluation will be discussed. All participants are invited to communicate their own experience or questions during the duration of this workshop.
Fluorescence Microscopy-Workshop (Olympus)
"Photoconvertible proteins as a useful tool for high-throughput, confocal and TIRF microscopy"Speaker 1: Simone Baltrusch (University of Rostock)
Proteins changing upon UV irradiation their fluorescence to a longer wavelength are called photoconvertible proteins. In contrast to the photoactivatable protein PA-GFP, which is colourless before activation, these proteins are visible from the outset. In live cell imaging photoconvertible proteins are very useful and thus, will be introduced in this workshop. In detail, analysis of protein mobility and identification of immobile fractions will be shown by the use of photoconversion and compared to fluorescence recovery after photobleaching (FRAP). Furthermore a photoconversion based high-throughput approach to determine the intracellular protein lifespan will be introduced. Finally the applicability of the photoconvertible protein Dendra2 in total internal reflection fluorescence (TIRF) microscopy will be demonstrated. All methods will be discussed in detail with the participants.
"The new Cell^R-TIRF-advanced methods in TIRF microscopy"
Speaker 2: : Helge Schmidt (Olympus GmbH)
Protein Tracking-Workshop
“Selective tracking of proteins and cells using Photoactivatable Green Cherry”Speaker 1: Arkadiusz Welman (University of Edinburgh)
Fluorescent highlighter proteins including photoactivatable fluorescent proteins and photoconvertible fluorescent proteins are powerful tools in modern cell biology, but they have also some limitations. In order to overcome some of these limitations a new fluorescent highlighter protein, Photoactivatable Green Cherry (GPAC), has been recently developed. GPAC displays unique “continuously red – photoactivatable green” two color fluorescence properties. It enables real-time tracking of selected subpopulations of proteins and organelles in the cell or of cells within tissues and whole organisms, with constant reference to the entire population of the probe. The advantages of GPAC in live cell imaging will be demonstrated using a few biological examples. The participants are invited to discuss the applicability of the GPAC probe to their own research.
“SNAP-tag Technology: Multiplex Tagging Tools for the Study of Protein Dynamics in Living Cells and beyond”
Speaker 2: Kristin Schnettler (New England Biolabs GmbH)
The creation of cell lines expressing proteins fused to intrinsically fluorescent tags (e.g., GFP) and their use in studies of protein localization has become routine. However, the fluorescent signal from such tags cannot be routinely switched on at will, precluding some time-resolved studies of expression, localization and degradation. A complementary and more flexible imaging system based on a family of small, self-labeling protein tags has recently been introduced by New England Biolabs. SNAP-tag and CLIP-tag, orthogonal tags with a variety of fluorescent substrates, are suitable for imaging intracellular and cell surface proteins in live cells. These tags can also be used for time-resolved dual labeling, highly efficient covalent pull-downs and other biochemical assays. Furthermore, a collection of non-fluorescent substrates that block SNAP and CLIP-tag reactivity enable pulse-chase studies and an examination of the temporal dynamics of nascent protein synthesis and complex formation in live cells. In this workshop, several studies of protein localization, turnover and receptor internalization using SNAP- and CLIP-tags in living cells are demonstrated.