Technische Universität Wien
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1D/2D gel electrophoresis

 

Electrophoresis is based on mobility separation of charged particles in liquid medium with an applied electrical field. The ion velocity is proportional to the electrical field and the charge state, whereas it is inverse proportional to the analyte radius and suspension viscosity. Analytes of interest are proteins, amino acids and intact cells.

Within proteomics, biological samples (celllysate, serum, blood ...) are separated after individual preparation and separated based on their isoelectric point (pI) or molecular weight (MW). Proteins can be characterized based on those parameters afterwards either in one singular (1D) or combined approach (2D) as well as Western Blot approaches. 

The protein identification itself, is based on mass spectrometry using peptide mass fingerprint (enzymatic cleavage pattern) and/or peptide sequencing (MSMS pattern).

FLA 9500

Typhoon FLA 9500 is a versatile laser scanner for biomolecular imaging applications including sensitive and quantitative measurements of radioisotopic labels by storage phosphor, chemifluorescent Western blots, and multiplex fluorescence as well as digitization of colorimetric stains (e.g., Coomassie Blue and silver-stained gels). The system supports both 2-D Difference Gel Electrophoresis (DIGE) and Amersham Western blotting systems.

 

HCT ion trap online coupled to an Ultimate 3000 (nL System) 

The HTCplus is also an ion trap mass spectrometer (IT-MS) which is by default equipped with an orthogonal electrospray ion trap (ESI). Compared to the Esquire 3000plus this instrument is characterized with a better sensitivity, a greater dynamic range and a faster scan speed.

In our lab this instrument is operated with a pulsed dynamic focusing (PDF)-MALDI ion source. This ion source works under atmospheric pressure and is, therefore, a useful supplement to high-vacuum MALDI.