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Two-dimensional electrophoresis in polyacrylamide gels

The proteomics unit of the University of Valencia has the following equipment for the separation and analysis of proteins by two-dimensional electrophoresis:

Ettan IPGPHOR II IEF: isoelectric focusing system to carry out the separation of proteins in a first dimension (pl), GE Healthcare.

Ettan DALTsix y Hoefer SE 600 Ruby: vertical electrophoresis systems for the separation of proteins in a second dimension (PM), GE Healthcare.

ImageScanner II: scanner for visible densitometry of revealed gels with conventional stains (silver, coomassie), GE Healthcare.

ImageMaster 2D Platinum: image analysis and differential expression software for the analysis of revealed gels with conventional stains, GE Healthcare.

Typhoon™ 9400: high-resolution scanner for the reading of fluorescence, chemiluminescence and radioisotopes which allows images of gels roots where proteins with fluorescent or chemiluminiescence mark (DIGE technology), GE Healthcare have been separated. 

DeCyder v6.5™: software for the analysis of differential expression of DIGE gels, GE Healthcare.

EDA v1.0: (Extended Data Analysis) software which is presented as an additional analysis module to Decyder which allows carry out an exhaustive analysis of the data, so it can analyse automatically the samples to be studied as to analysis of components, expression profile and discriminant analysis, GE Healthcare.

Analysis by mass spectrometry: molecular analysis, identification, sequencing and characterisation of proteins.

MALDI TOF-TOF 5800 ABSCiex: mass spectrometry with MALDI ionisation source and analszer of flight time. This mass spectrometer allows to carry out the following analysis:

  • Molecular weight determination of proteins, peptides and biomolecules.
  • Identification and characterisation of peptides and proteins.
  • Obtaining of molecular imaging of tissues.

QTRAP 5500 ABSCiex: mass spectrometer with ESI ionisation source and hybrid triple quadrupole analyser, linear ion trap. This mass spectrometer allows to carry out identifications of peptides and proteins and it is essential in the analysis of quantitative proteomics managed by SRM and MRM.

Triple Tof 5600 ABSCiex: mass spectrometer with ESI ionisation source and hybrid double quadrupole analyser, flight time. This mass spectrometer allows to carry out the following analysis:

  • Identification and characterisation of peptides and proteins.
  • Characterisation of post-translational modifications.
  • Differential expression of complex protein samples: identification of biomarkers. (iTRAQ, label free).
 
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