Yeast and Human Protein Extracts for Mass Spectrometry Method Development and Instrument Validation Poster

Part # PS187

Abstract

Sergei Saveliev, Ethan Strauss, Mike Rosenblatt and Marjeta Urh
Promega Corporation, Madison, WI

To test protein mass spec sample preparation procedures and validate mass spec instrument performance, complex biological samples are required. Total cell protein extracts provide this desired sample complexity. However, to be compatible with mass spec applications, such extracts should meet a number of design requirements:

• Compatibility with LC/MS (free of detergents, etc.)
• High protein integrity (minimal level of protein degradation and non-biological PTMs)
• Compatibility with common sample preparation methods such as proteolysis, PTM enrichment and mass-tag labeling
• Lot-to-lot reproducibility

Here we describe total protein extracts from yeast and human cells that meet the above criteria. Two extract formats have been developed:

• Intact protein extracts; primary use - sample preparation method development and optimization
• Pre-digested extracts (peptides); primary use - instrument validation and performance monitoring

 

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