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Protocols and Applications

Protocol

High throughput 4D-Proteomics – Application of dia-PASEF® and the Evosep One for short gradients

In this application note, we demonstrate the benefits of dia-PASEF technology on the timsTOF Pro platform coupled to an Evosep One for high-throughput in-depth proteome analysis of up to 300 samples per day (SPD). We quantify about 5200 protein groups in only 21-minutes run time. In 4.8-minutes ultr... More


Protocol

Elucidation of metabolic changes in HFD-ApoE–/– model by SP6 peptide: A flow injection analysis magnetic resonance mass spectrometry (FIA-MRMS) study

The employment of natural peptides in the treatment of cardiovascular diseases is an attractive option to pharmacological therapy. In this study, a novel antihypertensive peptide from Spirulina Platensis (SP6) was employed to evaluate its potential anti-atherosclerotic effect in high fat diet ApoE–/... More


Protocol

Magnetic Resonance Mass Spectrometry (MRMS) discriminates yeast mutants through metabolomics

A eukaryote model of Saccharomyces cerevisiae was used to study the methylglyoxal pathway by Magnetic resonance mass spectrometry (MRMS). It was discovered that glutathione plays a major role in driving metabolomic differences between different strains. It is expected that metabolomics-based discrim... More


Protocol

Proteomic Interrogation of Primary Insulin-secreting Pancreatic β-cells

In this work we leveraged the analytical performance of the timsTOF Pro to identify expression of low-level enzymes for the first time in primary insulinsecreting beta cells (β-cells). We found that these proteins are critical, functional components of key metabolic pathways which can support β-cell... More


Protocol

Studying protein-drug complexes under native conditions in the low nM range using MRMS

Protein ligand complexes need to be preserved through the electrospray and detection process. This work describes high resolution native mass spectrometric measurements of the enzyme carbonic anhydrase II (Figure 1) and the non-covalent complex with the substrate acetazolamide (Figure 2) using a sci... More


Protocol

Incorporating CCS values to enable 4-dimensional annotation of metabolic features

Reliable annotation of metabolites for LC-MS/MS based data requires the adept combination of many parameters such as retention time and accurate mass. To extend our parameter portfolio, we analyzed the high reproducible measured CCS values using trapped ion mobility spectrometry (TIMS) to provide ad... More


Protocol

prm-PASEF®: enabling high-throughput, high sensitivity targeted proteomics

In comparison with standard selected and parallel reaction monitoring (SRM and PRM), the prm-PASEF increases the number of peptides that can be targeted in a single acquisition method, without compromising the selectivity or the sensitivity. Using a prototype acquisition software, we targeted 201 is... More


Protocol

MRMS powered single cell metabolomics – Quantification of picogram amounts of a biocatalytic product from few living cells

Probing the metabolic state of living microbial cells via their secreted metabolic products is highly challenging as it requires measures to maintain the cellular integrity and to collect the analytes. This application note demonstrates how direct infusion magnetic resonance mass spectrometry (DI-MR... More


Protocol

Ultra-high sensitivity proteomics on the timsTOF SCP

The timsTOF Pro platform, introduced in 2017, already featured a sensitivity boost from the Parallel Accumulation SErial Fragmentation (PASEF) technology, which provides time and space focusing of the ions in the Trapped Ion Mobility Spectrometry (TIMS) tunnel. The timsTOF SCP platform further enhan... More


Protocol

Rapid and sensitive label-free proteome quantification on the timsTOF Pro 2

Shotgun proteomics capacity to deliver in-depth information on the identification, the relative amount, and the modification patterns of ever larger fraction of the proteins present in a sample has made it an essential approach in most biological research areas. The recent introduction of Parallel A... More