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Cell lysis is the first step when extracting proteins for mass spectrometry applications. Fractionation may be required to enrich for proteins of interest or to study protein localization. The kits featured here are convenient, easy to use, and have been validated for downstream analysis with Thermo Scientific mass spectrometers. They include options for either total protein extraction or isolation of proteins from specific cellular fractions. The Thermo Scientific EasyPep Mini MS Sample Prep Kit provides pre-formulated reagents and a robust method for the preparation of high quality samples that are ready for MS analysis in less than 3 hours.
Use this rapid mass spec sample prep kit for protein extraction, digestion and peptide clean-up in 2-4 hours.
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Thermo Scientific EasyPep Mini MS Sample Prep Kit | Mem-PER Plus Membrane Protein Extraction Kit | Subcellular Protein Fractionation Kit for Cultured Cells | |
Fractions isolated | Primarily cytosolic | Integral and membrane-associated proteins | Nucleus, cytoplasm, membrane, cytoskeletal, chromatin-bound |
Amount of sample processed | 20 samples of 10-100 µg of cell pellet or tissue | 50 samples of 5 million cells each | 50 samples of 2 million cells each (20 µL packed) |
Sample processing time (pre-digestion) | <30 min | 1 hr | 2–3 hr |
Digestion proteases included? | Yes | No | No |
Total sample preparation time | 1.5-3.5 hr | 24-48 hr | 24-48 hr |
Representative quantitation of protein enrichment of marker proteins (Hsp90 and histone 3) using TMTsixplex reagents in different subcellular fractions relative to whole cell lysate. The red bars show the relative intensity of TMT reagent reporter ions for unique marker proteins. Subcellular fractions were prepared using the Thermo Scientific Subcellular Protein Fractionation Kit for Cultured Cells (Cat. No. 78840).
Identification of peptides by LC-MS/MS from immunoprecipitated Na+/ K+-ATPase transport complex extracted with the Mem-PER Plus Kit (Cat. No. 89842). Samples were processed by in-solution digestion according to the kit protocol, and analyzed on the Orbitrap Fusion mass spectrometer.
Protein | Peptide sequence | Xcorr value |
---|---|---|
Na+/K+-transporting ATPase subunit beta-1 | VAPPGLTQIPQIQK | 2.67 |
VGNVEYFGLGNSPGFPLQYYPYYGK | 4.81 | |
AYGENIGYSEK | 2.52 | |
SYEAYVLNIVR | 3.17 | |
Na+/K+-transporting ATPase subunit alpha-1 | LSLDELHR | 2.20 |
QGAIVAVTGDGVNDSPALK | 4.03 | |
NIAFFSTNCVEGTAR | 3.29 | |
SPDFTNENPLETR | 4.46 | |
TSATWLALSR | 3.18 | |
AVFQANQENLPILK | 2.01 |
For Research Use Only. Not for use in diagnostic procedures.