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  • INTERFEROMETRY

    Pall ForteBio site [(http://www.fortebio.com/octet-specs.html). ] The OCTET RED 96 device is a practical alternative to surface plasmon resonance devices. Based on bio-layer interferometry, it is useful for our quantitative measurements. By using dedicated biosensors (8 in a row), it provides kinetics rate constants (Kon, Koff) useful to characterize protein/protein and Ag/Ab interactions, in 96-well (...)

  • BRET and HTRF

    Bioluminescence Resonance energy transfer) et HTRF (Homogenous Time-Resolved Fluore

    The BIOS group is using a Mithras 2 LB243 multimode plate reader equipped with a monochromateur and appropriate filters, for BRET, BRET2, FRET and HTRF data, in populations of living cells.We also use a Tristar 2 LB942 S. By these means, we measure β-arrestin recruitment to GPCRs, G protein trimer dissociation, GPCR dimerization and internalization, second messenger production, antibody/ receptor recognition, etc. Principle of the BRET-based G protein assay (upper image). G protein (...)

  • RPPA

    Reverse-Phase Protein Array

    In 2009, we developed in-house reverse-phase protein arrays (RPPA), using highly sensitive infrared detection. This technology allows the detection of phosphorylated proteins in the sub-attomolar range, from minute amounts of biological material. A phosphorylated protein of interest can then be quantified on-slide, from multiple experimental conditions, paving the way for high-throughput quantification of gonadotropin-induced signalling pathways. Dupuy et al., Proteomics (2009) 9, (...)

  • NANOPRO

    ProteinSimple site

    By the end of 2011, together with the PRC Unit, the BIOS group acquired the Nanopro technology (Protein Simple). This technology is based on isoelectric focusing in capillaries, to separate post-translationally modified proteins and quantify the various isoforms, from minute amounts of biological samples. Detection of phosphorylated ERK from untreated (fushia) or FSH-stimulated (blue) HEK293 cells stably expressing the FSH-R. PI values are plotted against signal intensities. Different (...)

  • FRET

    Fluorescence Resonance Energy Transfer

    Intramolecular FRET has been successfully implemented in our group, to monitor the production of second messengers or the activation of kinases in living cells, upon hormonal stimulation. So far, we are able to measure the production of *cAMP, *DAG and *Ca2+ and the activity of *ERK MAP kinases, *PLC and PKC, **PKA kindly provided by *Dr Jin Zhang (John Hopkins University, Baltimore, USA) and by **Pr Michiyuki Matsuda (Kyoto University, Japan). PKA activity assayed by FRET in HEK293 cells (...)

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