ArticleScientific reports2018
Hit-Gel: Streamlining in-gel protein digestion for high-throughput proteomics experiments.
Article in Scientific reports, 2018. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 10 papers.
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10 citing papers in PubMed.
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- Crop Proteomics under Abiotic Stress: From Data to Insights.Plants (Basel, Switzerland) · 2022Review
- Topology of the redox network during induction of photosynthesis as revealed by time-resolved proteomics in tobacco.Science advances · 2021Article
- Spatially Enriched Paralog Rearrangements Argue Functionally Diverse Ribosomes Arise during Cold Acclimation in Arabidopsis.International journal of molecular sciences · 2021Article
- A Semiautomated Paramagnetic Bead-Based Platform for Isobaric Tag Sample Preparation.Journal of the American Society for Mass Spectrometry · 2021Article
- Arabidopsis REI-LIKE proteins activate ribosome biogenesis during cold acclimation.Scientific reports · 2021Article
- Bringing New Methods to the Seed Proteomics Platform: Challenges and Perspectives.International journal of molecular sciences · 2020Review
- Protein Complex Identification and quantitative complexome by CN-PAGE.Scientific reports · 2019Article
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5 authors.
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Abstract
In-gel digestion has been used as a standard method for the preparation of protein samples for mass spectrometry analysis for over 25 years. Traditional in gel-digestion procedures require extensive sample handling, are prone to contamination and not compatible with high-throughput sample preparation. To address these shortcomings, we have modified the conventional in-gel digestion procedure for high-throughput proteomics studies. The modified method, termed "High Throughput in Gel digestion" (HiT-Gel), is based on a 96-well plate format which results in a drastic reduction in labour intensity and sample handling. Direct comparison revealed that HiT-Gel reduces technical variation and significantly decreases sample contamination over the conventional in-gel digestion method. HiT-Gel also produced superior results when a single protein band was excised from a gel and processed by in-gel digestion. Moreover, we applied Hit-Gel for a mass spectrometry analysis of Arabidopsis thaliana protein complexes separated by native PAGE in 24 fractions and four biological replicates. We show that the high throughput capacity of HiT-Gel facilitates large scale studies with high sample replication or detailed fractionation. Our method can easily be implemented as it does not require specialised laboratory equipment.
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