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GEN Protocols

OMICS

OMICS

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


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