High Performance Liquid Chromatography (HPLC)

Application: Protein characterization (molecular weight, oligomeric state, protein fold)

Instrument: Dionex UltiMate 3000 (Thermo Scientific)

Location: VBC6, room 124

High-Performance Liquid Chromatography (HPLC) coupled with Size-Exclusion Chromatography (SEC) is a powerful method for determining protein molecular weight and quality. SEC separates proteins by size, with larger ones eluting first and smaller ones retained longer. Calibration with molecular weight standards allows estimation of protein size, but this applies mainly to globular proteins. Non-globular proteins, like elongated or unfolded ones, have larger hydrodynamic radii and may not follow the standard correlation.

Our facility is equipped with an HPLC system optimised for small-volume sample analysis (about 10 µL). The setup allows precise separation and detection, with the option to collect elution fractions.

To use this system, users must first complete mandatory training.
Measurements can be performed by ProTech staff upon request.