2D gels at Riederer's laboratory

Proteomics Satellite Service at the Centre de neurosciences psychiatriques (CNP), Cery | Poster describing the IR-labelling technology | References
 

Proteomics Satellite Service at the Centre de neurosciences psychiatriques (CNP), Cery

The laboratory of neuroproteomics at the CNP is led by Dr. Beat Riederer. This team is actively developing techniques for differential labeling and co-migration of several samples on one gel, coupled to very sensitive infrared detection or DIGE. Another focus of research is the labeling and characterization of proteins with reactive cysteines. For more info on their technologies or inquiry about possible collaborations, visit the Riederer's group web page (http://www.chuv.ch/psychiatrie/dp_home/dp-recherche/dp-recherche-centres/dp-cnp/dp-recherche-proteomique.htm)

 

  • Over 30 years of experience in electrophoresis and 2D gels (equilibrium and non-equilibrium IEF gels) (refs 1-7)
     
  • labelling of individual protein samples for infrared detection (IR680 and IR780 maleimides) allowing high-sensitivity multiplexing
     
  • 2D gels, mini-, midi-, maxi (7cm, 11cm, 18cm), Western blots, Coomassie blue, silver stain and DIGE
     
  • Differential IR-labelling of non-oxized and oxidized proteins in the same protein sample.
     
  • 2D gel analysis with Melanie 4 / Image master

oxi_non-oxi.jpg

Protein sample from a fibroblast cell culture were first labeled at thiol sites with DY680 maleimide (red), the proteins that were masked by oxidation and not available for the first labeling were reduced and so made available for the second step, a labeling with DY780 maleimide (green). The protein sample (5µg) was focused on a pH3-10 IEF strip in the first dimension and 12.5% SDS-PAGE in the second dimension. Down to 1 femto gramme of protein can be detected by the Odyssey Infrared Imaging System.

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Poster describing the IR-labelling technology

IRdyesRiederer.pdf  (4602 Ko)

References

Antonsson, B., D. Kassel, G. DiPaolo, R. Lutjens, B.M. Riederer, and G. Grenningloh, 1998, "Identification of in vitro phosphorylation sites in the growth cone-associated protein SCG10: effect of phosphorylation site mutants on microtubule-destabilizing activity". J. Biol. Chem. 273: 8439-8446.

Genoud, S., W. Pralong, B.M. Riederer, L. Eder, S. Catsicas, and D. Müller, 1999, " Activity-dependent phosphorylation of SNAP-25 in hippocampal organotypic cultures". J. Neurochem., 72: 1699-1706.

Riederer B.M., M.M. Shaw, 2002, "Sample application techniques for brain tissue in two-dimensional gel electrophoresis" In: Proceedings of the Swiss Proteomics Society, Eds. P.M.Palagi, M.Quadroni, J.S.Rossier, J.-C. Sanchez, R.Stöcklin. Fontis Media, Lausanne, pp. 232-235.

Riederer I.M., P. Robert, R.Porchet, J. Eyer, and B.M. Riederer, 2003, "Selective changes in the neuronal cytoskeleton of NF-H/LacZ mice". J.Neurosci. Res., 71: 196-207.

Porchet R., A. Probst, C. Bouras, E. Dráberova , P. Dráber, and B.M. Riederer, 2003, "Analysis of gial acidic fibrillary protein in the human entorhinal cortex during aging and in Alzheimer's disease". Proteomics, 3: 1476-1485.

Shaw M., and B.M. Riederer, 2003, "Sample preparation for two-dimensional gel electrophoresis". Proteomics, 3: 1408-1417.

Cueille N., Riederer I.M., Tallichet Blanc C., Chevalley S., Pfulg C., LoPresti L., and Riederer B.M., 2004, " MAP1B is a crosslinker of several proteins during axonal development." In: Proceedings of the Swiss Proteomics Society, Eds: P.M.Palagi, M.Quadroni, J.S. Rossier, J.-C. Sanchez, R.Stöcklin. Fontis Media, Bern, P17, pp.108-111.

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