Easy Proteomics Sample Preparation: Technical Repeatability and Workflow Optimization Across 8 Biological Matrices in a New Core Facility Setting.

ABSTRACT: Bottom-up proteomics relies on efficient and repeatable sample preparation for accurate protein identification and precise quantification. This study evaluates the performance of adapted SPEED (Sample Preparation by Easy Extraction and Digestion) protocol, a simplified, detergent-free approach tailored for various biological matrices, including lysis-resistant samples. Protein extraction and denaturation steps were refined for 8 biological matrices enabling standardized, cheap, and scalable proteomics analysis on 96-well plates. For tissue samples requiring downstream applications like Western blotting, we used a low-detergent RIPA buffer. Notably, the protocols demonstrate remarkable down-scalability, enabling robust proteomics measurements from as few as 3000 cells per sample for prepara

SUBMITTER: Karousi P 

PROVIDER: S-EPMC12576925 | biostudies-literature | 2025 Oct

REPOSITORIES: biostudies-literature