Generate stable isotope-labeled protein standards for quantitative mass spectrometry

Stable isotope-labeled peptides are routinely used as internal standards for mass spectrometry (MS) quantitation of enzymatically digested protein samples. However, stable isotope-labeled proteins are considered better standards because they control for MS sample-preparation loss and digestion variations. In this Product Article about generating MS standards, our scientists describe using in vitro translation (IVT) as an alternative to in vivo expression systems. Most IVT systems use bacteria or rabbit reticulocyte cell extracts; however, such systems have multiple limitations. In this study, researchers used an IVT cellular-extract system and expressed stable isotope-labeled protein standards effective for quantitative MS.

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Cell-free protein expression system for generating stable isotope-labeled proteins