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Promega Corporation

Mass Spec-Compatible Extracts

protein image

Our mass spectrometry-compatible yeast and human protein extracts are designed specifically for mass spec applications. The extracts are predigested and cleaned up by solid-phase extraction for immediate use in liquid chromatography/mass spectrometry (LC-MS) analysis. The yeast and human extracts are also offered in an intact undigested form to provide a test material for optimizing mass spec protein sample preparation. Extract reproducibility is assured by stringent control over the manufacturing process. Each protein extract is monitored by LC-MS to confirm compositional and protein integrity.

LC-MS Performance Monitoring

Promega extracts provide complex protein reference material that offers consistent peptide retention time, signal intensity and other critical performance parameters.

Mass spec profiles from 3 consecutive weeks.

Consistent mass spec profile of complex protein extracts. Each run used 1µg of human predigested protein extract injected into the instrument (Waters NanoAquity HPLC system interfaced to a Thermo Fisher Q Exactive™ Hybrid Quadrupole-Orbitrap Mass Spectrometer). Peptides were resolved with 2-hour gradient. Weekly monitoring with the human extract ensured consistent analytical performance of the instrument.

Model System for Method Development

These protein extracts provide ideal model systems for development and optimization of various sample prep methods, including isobaric tagging and enrichment.

Bar graph showing relative abundance tagged extracts.

Example of method development, isobaric tagging. Three aliquots of the human K562 predigested extract were labeled with three different TMT isobaric mass tag reagents (TMT126, TMT127 and TMT128) and mixed at a 10:5:1 ratio. Relative quantitative ratio between the mass tags was determined with an LTQ Orbitrap Velos Pro™ mass spectrometer. The data show good correlation between the expected and observed protein quantitation levels.

Reliable and Reproducible Data

Lot-to-lot consistency of extracts is monitored by various protein and peptide qualitative and quantitative methods, including LC-MS and amino acid analysis. Our manufacturing process assures compatibility with LC-MS by monitoring nonspecific protein fragmentation, non-biological post-translational modifications and, for digested extracts, minimal undigested peptides.

QC specifications for MS Compatible Human Protein Extract, Digest.

Mass spec profiles from 3 consecutive weeks.

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