- Protein separation and image analysis.
Extraction of proteins for 2D-DIGE (Two-Dimensional Difference Gel Electrophoresis) analysis. Analysis of the differential expression of proteins using 2D-DIGE. Digestion of the differentially-expressed proteins with trypsin.
- Mass spectrometry analysis and protein identification.
Identification of the proteins expressed differentially in 2D-DIGE or in samples of other origin, using liquid chromatography (nano-HPLC) and ion trap mass spectrometry.
Instituto de Investigación Sanitaria del Principado de Asturias (ISPA). Edificio FINBA
Avenida Hospital Universitario s/n 33011 Oviedo Asturias, Spain.
Protein separation and image analysis.
- 2D-DIGE (Two-Dimensional Difference Gel Electrophoresis)
- First dimension: Ettan™ IPGphor™ 3 Isoelectric Focusing System (GE Healthcare Life Sciences)
- Second dimension: Ettan™ DALTsix Large Vertical System (GE Healthcare Life Sciences)
- Image analysis:
- Ettan™ DIGE Imager (GE Healthcare Life Sciences)
- Decyder Image Analysis Software (GE Healthcare Life Sciences)
Mass spectrometry analysis and protein identification (LC-MS/MS).
- Peptide separation:
- nano-HPLC (LC Packings, Ultimate model)
- LC Packings, FAMOS model)
- Mass Spectrometer - AB Sciex - API 2000 QTRAP
Specialist training is necessary to perform protein separation in two-dimensional gels and to identify differentially-expressed proteins, so all procedures will be carried out by the platform’s staff. Given the specificities of this service, it is necessary to contact a member of the platform staff if you would like to use it.
- Protein extraction, differential expression analysis and digestion with trypsin:
- €300 / 26x22 cm gel.
- €30 / hour of image analysis
- Mass spectrometry:
- Mass spectrometry analysis of differentially expressed proteins: €45 / sample.
- Identification of proteins from mass spectra in databases: €30 / hour.