Nejvíce citovaný článek - PubMed ID 33777071
SP3 Protocol for Proteomic Plant Sample Preparation Prior LC-MS/MS
The success of bottom-up proteomic analysis frequently depends on the efficient removal of contaminants from protein or peptide samples before LC-MS/MS. For a peptide clean-up workflow, single-pot solid-phase-enhanced peptide sample preparation on carboxylate-modified paramagnetic beads (termed SP2) was evaluated for sodium dodecyl sulfate or polyethylene glycol removal from Arabidopsis thaliana tryptic peptides. The robust and efficient 40-min SP2 protocol, tested for 10-ng, 250-ng, and 10-µg peptide samples, was proposed and benchmarked thoroughly against the ethyl acetate extraction protocol. The SP2 protocol on carboxylated magnetic beads proved to be the most robust approach, even for the simultaneous removal of massive sodium dodecyl sulfate (SDS) and polyethylene glycol (PEG) contaminations from AT peptide samples in respect of the LC-MS/MS data outperforming ethyl acetate extraction.
- Klíčová slova
- Arabidopsis thaliana, LC-MS/MS, SP2, detergent, ethyl acetate extraction, magnetic beads, peptide clean-up, polyethylene glycol, sodium dodecyl sulfate,
- MeSH
- chromatografie kapalinová metody MeSH
- dodecylsíran sodný MeSH
- kapalinová chromatografie-hmotnostní spektrometrie * MeSH
- peptidy analýza MeSH
- polyethylenglykoly * MeSH
- proteomika metody MeSH
- tandemová hmotnostní spektrometrie metody MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- dodecylsíran sodný MeSH
- ethyl acetate MeSH Prohlížeč
- peptidy MeSH
- polyethylenglykoly * MeSH