Simple gradient analysis of phosphopeptides

Instrument Type: HPLC

The Thermo Scientific HPLC system is applied for the analysis of phosphopeptides. The sample of ß-casein was reduced and carboxymethylated, and digested for 9 hours with immobilized trypsin. The sample was fractionated by immobilized metal affinity chromatography (IMAC) in ferric form, before analysis using a Thermo Scientific Acclaim 300 C18 column, with UV detection at 214 nm. Note pyrophosphate is a UV transparent chelating agent that removes or masks interfering metal ions. The chromatograms shown are: A – fraction retained by IMAC B – unretained fraction C – unfractionated peptides

Four high resolution peptide maps using a Thermo Scientific Acclaim PolarAdvantage II HPLC column

Instrument Type: HPLC

The Thermo Scientific UltiMate 3000 RSLC system is applied for the analysis of ovalbumin, fetuin, β-casein and bovine serum albumin. The separation is performed on a Thermo Scientific Acclaim PolarAdvantage II HPLC column, with UV detection at 210 nm.