Overview of specific training stations:
University of Cambridge |
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| Type of training available: | - We routinely make model bilayer membranes (giant unilamella vesicles), and we observe them by optical microscopy. Optical traps are used to move the vesicles, and deform them. This gives access to membrane mechanics, and can also allow processes such as membrane fusion to be studied. - We have a surface science lab, for the physical characterisation of liquid surfaces, including various forms of tensiometry and rheology. - We develop instrument automation, to perform long time optical microscopy (phase contrast; epi-fluorescence; confocal) experiments on cell cultures, including image analysis code for cell segmentation. |
| Number of students which can be hosted: | One student at a time can be hosted; probably up to 2 students per year. |
| Link: | http://www.bss.phy.cam.ac.uk/~pc245/ |
International Institute of Molecular and Cell Biology (IIMCB) Warsaw |
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| Type of training available: | Biochemical and functional assays in cultured mammalian cells: - RNAi techniques (e.g. siRNA transfection, production of esiRNA), - RNAi-based screens of endocytic proteins, - Transcriptional assays (luciferase-based reporters and qRT-PCR), - Biochemical assays (e.g. western blotting, cell fractionation, co-immunoprecipitation, GST pull-down), - Signaling assays (e.g. cell proliferation/survival, apoptosis, activation of various signaling pathways), - Protein localization studies by confocal microscopy, - Microscopy-based assays for endocytosis and cell proliferation, quantitative analysis of images (up to 4 channels), - xxx Molecular biology techniques (cloning, mutagenesis, etc).
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| Instruments available: | Cell culture equipment, including Amaxa nucleofection device 96-well plate luminometer RT-PCR cycler Odyssey Infrared Imaging System for quantitative western blotting 3 confocal microscopes Olympus Scan^R microscope for automated imaging in multiwell plates |
| Number of students which can be hosted: | Preferably 1 student for training at one time, maximum 2 students at the same time |
| Link: | http://www.iimcb.gov.pl/Miaczynska-Laboratory-research-focus.html |
Curie Institute |
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| Type of training available and Instruments available: | - Clathrin-independent endocytosis, intracellular/retrograde trafficking - Cellular biochemistry: quantitative trafficking assay, trafficking probes and tracers - Model membrane reconstitution of trafficking events - Glycosphingolipid synthesis and reconstitution in cells - Proteomics of clathrin-independent endocytosis and retrograde transport - Membrane mechanics, biology/physics interface - Trafficking and signaling - Nikkon Imaging center |
| Number of students which can be hosted: | 2-3 fellows |
| Link: | http://umr144.curie.fr/fr/equipes-de-recherche/trafic-signalisation-et-ciblage-intracellulaires-ludger-johannes/trafic-signali |
University of Geneva |
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| Type of training available and Instruments available: | We are specialists of lipid biophysics. We provide all techniques for making Small Unilamellar Vesicles (SUVs), Large Unilamellar Vesicles (LUVs) and Giant Unilamellar Vesicles (GUVs) :-spontaneous growth, sonication, extrusion and electroformation. We also have expertise in in vitro reconstitution assays with proteins and liposomes. Finally, we have expertise in Optical Tweezers, nanoforce measurements, and fast photonic imaging (confocal and other). Also any protein biochemistry technique (purification, FPLC, fluorescent labelling) are also available. |
| Number of students which can be hosted: | Maximum 2 fellows |
| Link: | http://cms.unige.ch/sciences/biochimie/-Aurelien-Roux-Lab- |
Weizmann Institute |
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| Type of training available and Instruments available: | Techniques offered at the training station of Gideon Schreiber / Nanaocha Sharma Protein-Protein interactions: Our laboratory has extensive expertise in measuring and analyzing protein-protein interactions, quantitative, qualitative and in vivo. Quantitative analysis in vitro: 1. Using SPR (we have a ProteonX36, that allows 36 protein-protein interactions to be measured simultaneously). This method measures on and off rates as well as binding affinities. For more information see Bravman T, Bronnera V, Nahshol O, Schreiber G (2008) Cellular and Molecular Bioengineering, 1, 216–228. 2. Stopped-flow measurements of protein interactions in solution. We have a stopped-flow instrument, and extensive experience in it. Rapid, semi-quantitative analysis using crude cell extracts. We have developed a method (FRETex) that takes advantage of the FRET signal between proteins to measure their interactions directly in small quantities of crude E.coli extracts. The method is very fast, and allows the parallel measurements of many different samples. For more details see: Khait R, Schreiber G (2012) Protein Eng Des Sel, Sep 25. Qualitative integration analysis in vivo: We use routinely a method termed proximity ligation assay (PLA) that allows for the detection of protein-protein interactions in cells without the need for any transfection. The method provides both semi-quantitative as well as spatial information. Details about the method can be found at: http://www.immunoportal.com/modules.php?name=News&file=article&sid=205 Protein-production. Our laboratory has extensive experience in protein-production. This includes production in many different vectors in E.coli, including refolding from inclusion bodies. In addition, we do protein-production in baculovirus (insect cells) using a simplified, fast method.
Real time PCR.Our laboratory is very experienced in quantitative rtPCR, including the use of very small sample volumes that allow the parallel handling of many different samples at a relatively short time |
| Number of students which can be hosted: | |
| Link: | http://www.immunoportal.com/modules.php?name=News&file=article&sid=205 |
Ruhr University, Bochum |
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| Type of training available and Instruments available: | Techniques offered at the training station of Rolf Heumann/Panos Athanasopoulos
Cell Biology: Our laboratory has extensive expertise in preparing primary cell cultures of the nervous system (astrocytes, neurons, oligodendrocytes) analyzing proteins by immunohistochemistry including some live cell imaging. Primary cultures are taken mainly from transgenic mice. We have access to transgenic mouse facilities. We also have animal and human induced pluripotent stem cells (iPS) cultures. Tools for investigating mechanisms of cellular apoptosis and Ras signaling pathways are available. Protein transduction: We have extensive experience with techniques to introduce proteins into the cell via protein cell membrane penetration mechanisms. Microscopy: The Ruhr University (RUBION) has recently established super resolution microscopy using STED (stimulated emission depletion) techniques. This service is open for TRANSPOL fellows. We have experience in measuring intracellular Ca2+ levels in live cells. Protein-production: We have experience in protein-production. This includes production in different vectors in E.coli. , protein-production in baculovirus (insect cells) and in transfected Hela cells. We produce transcription factor proteins used for cellular reprogramming. |
| Number of students which can be hosted: | One student at a time can be hosted |
| Link: | http://www.ruhr-uni-bochum.de/mol-neurobio/Lehrstuhl/Biochemie2en.htm |
Ruhr University, Bochum |
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| Type of training available and Instruments available: | Department of Biochemistry I - Receptor Biochemistry, Ruhr University Bochum, Bochum, Germany (Prof. Hollmann)
Heterologous expression and functional analysis of membrane proteins in Xenopus laevis oocytes: -- Subcloning of desired cDNAs into appropriate vector -- In vitro cRNA synthesis from cDNA construct -- Surgical harvesting of Xenopus oocytes and microinjection of cRNA -- Two-electrode voltage clamp measurements of currents through membrane proteins with integral ion channels -- Dose-response curves, I/V curves, ion permeability determination -- Oocyte membrane preparations and Western blots of membrane proteins -- Confocal imaging of fluorescence-labeled membrane proteins-- Interaction analysis between membrane proiteins in oocytes by FRET or BiFC |
| Instruments available: | -- State-of-the-art frog-keeping facility -- 3 Injection setups for microinjection into frog oocytes -- 4 Two-electrode voltage clamp set-ups -- 1 Leica TCS SP2 AOBS confocal microscope -- 1 Real-time PCR machine Roche LightCycler -- 1 Applied Biosystems 3130xl Genetic Analyzer |
| Number of students which can be hosted: | Preferentially 1 student at a time; maximum 2 students simultaneously |
| Link: | http://www.ruhr-uni-bochum.de/bc1/index_en.html |
| Type of training available: | Department of Physical Chemistry I, Ruhr University Bochum, Bochum, Germany (Prof. Christian Herrmann)
Biochemical and biophysical characterizations of protein-protein interactions. |
| Instruments available: | Instrumentation available: Micro-calorimetry: Isothermal titration calorimetry (ITC), Differential scanning calorimetry (DLS); Circular dichroism (CD) – spectroscopy; Fluorescence Spectroscopy; Fluorophore-based Stopped-Flow system and Temperature Jump system. |
| Number of students which can be hosted: | One student at a time can be hosted |
| Link: | http://www.ruhr-uni-bochum.de/proin/ |