ArticlePlant methods2018
Evaluation of sample preparation methods for mass spectrometry-based proteomic analysis of barley leaves.
Article in Plant methods, 2018. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 26 papers.
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Who cites it
26 citing papers in PubMed.
- Proteomics and Alkaloid Signature of Narrow Leaf Lupin Leaves Reveal Key Enzymes in Quinolizidine Alkaloid Biosynthesis and Transportation.Journal of agricultural and food chemistry · 2026Article
- Toward simple, rapid, and deep plant proteome analysis with an in-cell proteomics strategy.bioRxiv : the preprint server for biology · 2025Article
- Impact of Sample Preparation Strategies on the Quantitative Accuracy of Low-Abundance Serum Proteins in Shotgun Proteomics.Journal of proteome research · 2025Article
- Contamination of Wheat Flour and Processed Foodstuffs with Soybean and Mustard Allergenic Proteins.International journal of molecular sciences · 2025Article
- Identification of Microbial Strains via 2D Cross-Correlation of LC-MS Data.Journal of the American Society for Mass Spectrometry · 2024Article
- Review
- Differences in Protein Capture by SP3 and SP4 Demonstrate Mechanistic Insights of Proteomics Clean-up Techniques.bioRxiv : the preprint server for biology · 2024Article
- Getting Ready for Large-Scale Proteomics in Crop Plants.Nutrients · 2023Article
- Detergent-Assisted Protein Digestion-On the Way to Avoid the Key Bottleneck of Shotgun Bottom-Up Proteomics.International journal of molecular sciences · 2022Review
- Article
- SimPLIT: Simplified Sample Preparation for Large-Scale Isobaric Tagging Proteomics.Journal of proteome research · 2022Article
- Applications of Tandem Mass Spectrometry (MS/MS) in Protein Analysis for Biomedical Research.Molecules (Basel, Switzerland) · 2022Review
- Shotgun Proteomics as a Powerful Tool for the Study of the Proteomes of Plants, Their Pathogens, and Plant-Pathogen Interactions.Proteomes · 2022Review
- Does filter-aided sample preparation provide sufficient method linearity for quantitative plant shotgun proteomics?Frontiers in plant science · 2022Article
- The enigmatic clock of dinoflagellates, is it unique?Frontiers in microbiology · 2022Review
- Evaluation of Sample Preparation Strategies for Human Milk and Plasma Proteomics.Molecules (Basel, Switzerland) · 2021Article
- Reversible Click Chemistry Tag for Universal Proteome Sample Preparation for Top-Down and Bottom-Up Analysis.Journal of proteome research · 2021Article
- Application of Mass Spectrometry-Based Proteomics to Barley Research.Journal of agricultural and food chemistry · 2021Review
- Investigation of the Initial Host Response of Naïve Atlantic Salmon (Microorganisms · 2021Article
- SP3 Protocol for Proteomic Plant Sample Preparation Prior LC-MS/MS.Frontiers in plant science · 2021Article
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Authors and funding
5 authors.
Funding
No grant is acknowledged in the PubMed record.
Abstract
backgroundSample preparation is a critical process for proteomic studies. Many efficient and reproducible sample preparation methods have been developed for mass spectrometry-based proteomic analysis of human and animal tissues or cells, but no attempt has been made to evaluate these protocols for plants. We here present an LC-MS/MS-based proteomics study of barley leaf aimed at optimization of methods to achieve efficient and unbiased trypsin digestion of proteins prior to LC-MS/MS based sequencing and quantification of peptides. We evaluated two spin filter-aided sample preparation protocols using either sodium dodecyl-sulphate or sodium deoxycholate (SDC), and three in-solution digestion (ISD) protocols using SDC or trichloroacetic acid/acetone precipitation.
resultsThe proteomics workflow identified and quantified up to 1800 barley proteins based on sequencing of up to 6900 peptides per sample. The two spin filter-based protocols provided a 12-38% higher efficiency than the ISD protocols, including more proteins of low abundance. Among the ISD protocols, a simple one-step reduction and
conclusionsWe provide detailed protocols for efficient plant protein sample preparation for LC-MS/MS-based proteomics studies. Spin filter-based protocols are the most efficient for the preparation of leaf samples for MS-based proteomics. However, a simple protocol provides comparable results although with different peptide digestion profile.
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