Extended investigation of tube-gel sample preparation: a versatile and simple choice for high throughput quantitative proteomics

Scientific Reports
Leslie MullerChristine Carapito

Abstract

Sample preparation for quantitative proteomics is a crucial step to ensure the repeatability and the accuracy of the results. However, there is no universal method compatible with the wide variety of protein extraction buffers currently used. We have recently demonstrated the compatibility of tube-gel with SDS-based buffers and its efficiency for label-free quantitative proteomics by comparing it to stacking gel and liquid digestion. Here, we investigated the compatibility of tube-gel with alternatives to SDS-based buffers allowing notably the extraction of proteins in various pH conditions. We also explored the use of photopolymerization to extend the number of possibilities, as it is compatible with a wide range of pH and is non-oxidative. To achieve this goal, we compared six extraction buffers in combination with two polymerization conditions to further optimize the tube-gel protocol and evaluate its versatility. Identification and quantitative results demonstrated the compatibility of tube-gel with all tested conditions by overall raising quite comparable results. In conclusion, tube-gel is a versatile and simple sample preparation method for large-scale quantitative proteomics applications. Complete datasets are available...Continue Reading

References

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Nov 15, 2017·Nature Communications·Sophia DollMatthias Mann

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Citations

Jul 2, 2019·PLoS Computational Biology·Albert Tian ChenNikolai Slavov

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Datasets Mentioned

BETA
PXD008656

Methods Mentioned

BETA
PCA
scraping

Software Mentioned

factoextra
R
Studio
FactoMineR package
ProteomeXchange
FactoMineR
Mascot
[UNK]

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