This EVOSEP+ method is the ideal choice, when maximum proteome coverage in a single experiment is needed, especially when the amount of starting material is limited.
A long gradient with low overhead
The Extended method has a 88 min gradient, equivalent to a throughput of a little more than 15 samples per day, which is possible due to the short 4 min overhead between injections.
It runs with a gradient flow of 220 nl/min and is designed to be used with our EV-1106 column.
It takes advantage of our offset gradient strategy, which results in sharp peaks with highest possible peak capacity.
SIMILAR PERFORMANCE IN TWO DIFFERENT LABORATORIES
Reproducibility is a key aspect in clinical proteomics both in terms of limiting the technical variation between replica injections, but also the inevitable variation between instruments and laboratory procedures.
In a comparative pilot study in two different laboratories, similar proteome coverage is observed with around 35,000 unique peptides corresponding to 5000 proteins from 500 ng peptide load.
The high reproducibility comes from advanced calibration and flow control coupled with continuous self-monitoring and adjustments
A minor difference is observed at the lower loads.
MORE PROTEOME COVERAGE IN A SINGLE EXPERIMENT
Read the entire application note describing the method design, retention time reproducibility and the comparative pilot study.
Data courtesy: Professor Matthias Mann and Johannes Müller, Max-Planck Institute of Biochemistry, Proteomics and Signal Transduction on a Thermo Scientific Orbitrap Exploris 480 MS
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