Amarjeet Flora, Sergei Snovida, Ryan Bomgarden
Advances in mass spectrometry (MS) instrumentation has enabled routine analysis of complex protein samples. However, sample preparation methods are not standardized with many protocols taking 8-24 hours in addition to suffering from low peptide yields, poor digestion efficiency and low reproducibility. Here, we describe a simplified sample prep kit containing pre-formulated reagents and a standardized protocol that can be used to efficiently process 10μg to 100μg protein samples in less than 2 hours. In this study, we evaluated the scalability, compatibility, and reproducibility of this sample preparation kit compared to previous published methods. Our new standardized workflow yielded 10-20% higher number of peptides and proteins with lower missed cleavages (<90%) compared to other commercial MS sample prep kits and protein digest standards.
Thermo Fisher Scientific, Rockford, IL, USA, 61101