This document illustrates the effectiveness of the Agilent 3100 OFFGEL Fractionator for the prefractionation of complex protein or peptide mixtures. The tryptic digest of E. coli proteins was fractionated into 24 fractions, and peptides were identified using reversed phase liquid chromatography with mass spectrometric detection. The distribution of peptides and their isoelectric points in each fraction was analyzed. The results show efficient peptide fractionation and improved protein identification using the Agilent 3100 OFFGEL Fractionator.
This document describes an experiment where a tryptic digest of E. coli proteins was fractionated into 24 fractions using the Agilent 3100 OFFGEL Fractionator with IPG strips (pH 3-10). The technique utilizes OFFGEL electrophoresis to separate peptides based on their isoelectric point (pI) in a liquid phase, avoiding the tedious sample processing required by traditional gel-based methods.
The sample was focused with a maximum current of 50 pA.
The total run time, including column reconditioning, was 45 minutes.
A single well covers a pH interval of 0.26 units.