SP3 Protocol for Proteomic Plant Sample Preparation Prior LC-MS/MS
The result's identifiers
Result code in IS VaVaI
<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F00216224%3A14740%2F21%3A00121574" target="_blank" >RIV/00216224:14740/21:00121574 - isvavai.cz</a>
Result on the web
<a href="https://www.frontiersin.org/articles/10.3389/fpls.2021.635550/full" target="_blank" >https://www.frontiersin.org/articles/10.3389/fpls.2021.635550/full</a>
DOI - Digital Object Identifier
<a href="http://dx.doi.org/10.3389/fpls.2021.635550" target="_blank" >10.3389/fpls.2021.635550</a>
Alternative languages
Result language
angličtina
Original language name
SP3 Protocol for Proteomic Plant Sample Preparation Prior LC-MS/MS
Original language description
Quantitative protein extraction from biological samples, as well as contaminants removal before LC-MS/MS, is fundamental for the successful bottom-up proteomic analysis. Four sample preparation methods, including the filter-aided sample preparation (FASP), two single-pot solid-phase-enhanced sample preparations (SP3) on carboxylated or HILIC paramagnetic beads, and protein suspension trapping method (S-Trap) were evaluated for SDS removal and protein digestion from Arabidopsis thaliana (AT) lysate. Finally, the optimized carboxylated SP3 workflow was benchmarked closely against the routine FASP. Ultimately, LC-MS/MS analyses revealed that regarding the number of identifications, number of missed cleavages, proteome coverage, repeatability, reduction of handling time, and cost per assay, the SP3 on carboxylated magnetic particles proved to be the best alternative for SDS and other contaminants removal from plant sample lysate. A robust and efficient 2-h SP3 protocol for a wide range of protein input is presented, benefiting from no need to adjust the amount of beads, binding and rinsing conditions, or digestion parameters.
Czech name
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Czech description
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Classification
Type
J<sub>imp</sub> - Article in a specialist periodical, which is included in the Web of Science database
CEP classification
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OECD FORD branch
10611 - Plant sciences, botany
Result continuities
Project
Result was created during the realization of more than one project. More information in the Projects tab.
Continuities
P - Projekt vyzkumu a vyvoje financovany z verejnych zdroju (s odkazem do CEP)
Others
Publication year
2021
Confidentiality
S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů
Data specific for result type
Name of the periodical
Frontiers in Plant Science
ISSN
1664-462X
e-ISSN
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Volume of the periodical
12
Issue of the periodical within the volume
MAR
Country of publishing house
CH - SWITZERLAND
Number of pages
12
Pages from-to
„635550“
UT code for WoS article
000631851900001
EID of the result in the Scopus database
2-s2.0-85103216802