Proteomics-Protein Purification

The CBI Proteomics-Protein Purification “facility” allows to structure, on the whole site, the available resources dedicated to the study of proteins. It is organized around the resources in materials and personnel of the MCD and LMGM units. Its role is to offer to all the CBI staff the means to develop research projects which require protein purification and/or analysis by Mass Spectrometry.

CONTACTS

Low-pressure Chromatography :

      • Régine Capeyrou, IR-CNRS, MCD
      • Christophe Normand, IR-CNRS, MCD

Mass Spectroscopy :

      • Régine Capeyrou, IR-CNRS, MCD
      • Anne Caumont-Sarcos, IE-CNRS, LMGM

 

LOW-PRESSURE CHROMATOGRAPHY (FPLC)

For the low-pressure chromatography part, the persons in charge are Christophe Normand and Régine Capeyrou for the MCD unit and Violette Moralès for the LMGM. Their roles are to discuss upstream with the users the technical strategies that can be implemented with the material resources of the CBI, and then to ensure technical support and training of the users for an autonomous realization of the projects.

The pole is open to all CBI staff who are invited to contact the managers to discuss your projects

Available equipment

The CBI has 9 low and medium pressure liquid chromatography (FPLC) systems controlled by the UNICORN software. The MCD unit has 1 AKTA purifier and 1 AKTA basic and 1 AKTA pure in cold room and 2 AKTA Purifier at room temperature. The LMGM unit has 4 AKTA purifiers. These machines are not freely available and their use is subject to prior training. It is important to contact the manager, Christophe Normand (MCD) to discuss the implementation of your project.

FPLC system (MCD unit)

MASS SPECTROSCOPY

Proposed service: Identification of proteins in low complexity mixtures

Apparatus : MALDI AB Sciex TOF/TOF 5800 from IPBS (Hosting agreement with IPBS for use)

People from CBI authorized to use the MALDI :

Anne Caumont-Sarcos (IE, LMGM) and Régine Capeyrou (IR, MCD) have followed theoretical and practical training courses. They are in charge of setting up the mass spectrometry activity respectively at LMGM and MCD, accompanied by Cécile Albenne (MC, LMGM), mass spectrometry expert.

Principle : Excision of proteins from a denaturing gel colored with Coomassie blue, digestion of the sample with an adapted protease (mainly trypsin), analysis of the sample on the MALDI TOF/TOF, data processing.

Anne Caumont-Sarcos and Régine Capeyrou advise the users of LMGM or MCD before the analysis. They collect the bands excised on gel by the requesting team (see MALDI instructions and MALDI table). They prepare the samples and perform the analysis on MALDI TOF/TOF. They process the data obtained and report their results to the users.

A service including all the collected samples is proposed every 6 to 8 weeks.

The cost of the analysis is invoiced per sample to the requesting teams, taking into account the cost of the reagents for the preparation of the samples and the cost of using the MALDI TOF/TOF.

See also in «Technology»

Cytometry Drosophila transgenesis facility GenSCAN (Genomics Single Cell Analysis)