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Sample preparation remains one of the largest bottlenecks associated with mass spectrometry (MS) analysis, and consistent and reproducible sample preparation can make the difference between a successful analysis and a failed outcome. Thermo Scientific reagents and kits are optimized for sample preparation for every step of the process, from depletion strategies to reduce complexity, to protein extraction, enrichment, and clean-up for the isolation of low-abundance proteins. Post digestion, peptide enrichment, fractionation, and clean-up steps are often critical for proper analysis. The colorimetric and fluorometric Quantitative Peptide Assays enable sample normalization following digestion. AccelerOme: Automated sample preparation solution to simplify mass spectrometry sample preparationBreak free from the long reaction times, high costs, and method complexity of manual proteomic sample preparation with the Thermo Scientific AccelerOme automated sample preparation platform. With factory-supplied reagents and kits and step-by-step onscreen instructions, high reproducibility and confident results are delivered for both label-free and tandem mass tag (TMT) multiplexing strategies with minimal effort providing several reagents to perform Label-free, TMT 11-plex, and TMT pro16-plex workflows at different sample scales. The AccelerOme platform is optimized to fit into the Thermo Scientific Orbitrap mass spectrometer ecosystem, streamlining the entire workflow from sample preparation to data processing. EasyPep 96 Micro MS Sample Prep Kit and Peptide Clean-up PlateDesigned to simplify and standardize mass spectrometry sample preparation, whether you're working with 1 μg or up to 10 μg samples, the Thermo Scientific EasyPep Kit offers a comprehensive and user-friendly solution for mass spec sample prep on a large scale. With the Peptide Clean-up Plate, streamline peptide clean-up, embrace an efficient way of preparing your samples for downstream analysis. Key features of the Thermo Scientific EasyPep 96 Micro MS Sample Prep Kit Efficient and reproducible processing of cultured mammalian cells and tissues for proteomic mass spectrometry (MS) analysisOptimized to process protein samples from 1–10 μg with a high yield of MS-ready peptidesKit includes a peptide clean-up plate and solutions to prepare detergent-free peptide samples for direct LC-MS analysis or further sample processing such as isobaric tag (e.g., TMT reagent) labeling, and high pH reversed-phase fractionation with TMT/TMTpro pooled samples Key features of the Thermo Scientific Peptide Clean-Up Plate Efficient contaminant removal from 10 μg to 100 μg peptide samples following enzymatic digestionResin contained in each well provides excellent binding and recovery characteristics of peptide samples in preparation for mass spectrometry and other methodsPlate format allows the processing of multiple samples (1–96) in parallel in less than 30 minutesOrder now EasyPep Mini, Maxi, and 96-well plate MS Sample Prep Kits–simplified sample preparation solutions for mass spectrometrySample preparation of peptides for MS analysis is complex, with numerous steps and non-standard protocols, resulting in variable sample quality and poor reproducibility. To address these issues, the Thermo Scientific EasyPep MS Sample Prep kits have been designed using a standardized workflow to help improve reproducibility while also saving hands-on and processing time. HighlightsComplete—includes pre-formulated reagents for lysis through digestion, peptide cleanup columns, and an optimized protocol for processing up to 20 samples (mini-column), 8 samples (maxi-column), or one 96-well plateOptimized—streamlined protocol and reagents minimize the number of steps and time it takes to process samplesFlexible—reagents and protocol have been verified using cells, plasma, and tissue samples for 10 µg to 2 mg samplesTime-saving—sample processing has been reduced from more than 1 day to less than 4 hoursOrder now Click image to enlargeFigure 1. Comparison of sample preparation for MS analysis, using a traditional workflow and EasyPep MS Sample Prep Kits. Compared to a traditional workflow, the EasyPep MS Sample Prep Kit provide the same high-quality data in less time and with half the number of manipulation steps. Option for TMTpro labeling in workflows indicated. The number of steps and time have been reduced through the addition of Thermo Scientific Pierce Universal Nuclease as a replacement for sonication to reduce viscosity from nucleic acids, a rapid “one pot” reduction/alkylation solution for cysteine modification (carbamidomethylation, +57.02), and a trypsin/Lys-C protease mix for more complete digestion. In addition, these kits include peptide cleanup columns (or plates) and buffers to prepare detergent-free peptide samples for MS analysis in less than 3 hours. Compared to other methods and commercial kits, the EasyPep MS Mini Sample Prep kit produces MS ready peptides with higher protein yields and more unique protein group identifications. This is due to the high quality of the samples which have fewer contaminants and higher digestion efficiency compared to traditional overnight digestion with trypsin alone. Click image to enlargeFigure 2. HeLa cell lysates prepared using the EasyPep Mini MS Sample Prep Kit compared to other commercially available kits. The EasyPep Mini MS Sample Prep Kit provides similar reduction and alkylation efficiencies with more protein identifications and better digestion efficiency (higher percentage of zero missed cleavages) than other methods per microgram of sample analyzed. Thermo Scientific Pierce HeLa Protein Digest Standard (Cat. No. 88328) was used as a control sample. The EasyPep MS Sample Prep kits are available in three formats to support a wide range of sample numbers and input amounts. The EasyPep Mini and 96 MS Sample Prep kits are optimized to efficiently process protein samples of 10–100 µg with a high yield of MS-ready peptides. The EasyPep Maxi MS Sample Prep kit is optimized to process eight 0.5–2 mg protein samples ideal for phosphopeptide enrichment and other applications that require larger starting sample amounts. Alternatively, each Peptide Clean-Up Maxi Column can be used to process a combined set of isobaric tag-labeled samples (10–100 ug each, ≤2 mg total) for multiplex proteomic quantitation. Click image to enlargeFigure 3. The EasyPep MS Sample Prep kits use the same chemistry and standardized method to process plasma, cells, and tissues for downstream MS-based proteomic applications. Mini, 96-well, and maxi formats can be used to generate samples for label-free quantitation, SureQuant targeted peptide assays, TMT reagent labeling, or phosphopeptide enrichment. Easy-Pep Sample Prep kit selection guide EasyPep Mini Sample Prep KitEasyPep Maxi Sample Prep KitEasyPep 96 MS Sample Prep KitAccelerOme Sample Prep KitsAccelerOme TMT11plex KitsAccelerOme TMTpro KitsCat No.A40006A45734A45733A50944 and A50945A50946, A50947, A50948A50949, A50950, A50951Multiplexing None11plex16plexFormatColumnColumnPlate Quantity20 reactions8 reactions96 reactions Sample number 16,3611,3316,32Sample typeCells, tissue, plasma, serumCells, tissue, plasma, serumCells, tissue, plasma, serumcells, tissue, plasma, purified proteinsCapacity10–100 µg0.5–2 mg 10–100 µg10-100ug10-30ug, 30-100ugProcess combined, multiplex set of isobaric tag-labeled samplesNot recommendedYes (10–100 ug each)Not recommended Pooling and clean-up performed on AccelerOme System How-to-use video for EasyPep Mini MS Sample Prep kitUse this rapid mass spec sample prep kit for protein extraction, digestion, and peptide clean-up in 2-4 hours. Choose the right mass spec sample prep products for your sample type Cultured cellsSerum, plasma, or biofluidsTissuePurified proteinIntact proteinLysis/extraction✓ ✓ Abundant protein depletion✓ Optional Protein enrichment✓ ✓ ✓ ✓ ✓ Protein interaction crosslinking✓ ✓ ✓ ✓ ✓ Protein quantitation assays✓ Optional✓ Optional✓ Gel separationOptional Optional✓ Protein digestion✓ ✓ ✓ ✓ OptionalPeptide enrichment/fractionation✓ ✓ ✓ Peptide cleanup✓ ✓ ✓ ✓ OptionalDetergent removal✓ ✓ ✓ ✓ OptionalPeptide quantitation assays✓ ✓ ✓ ✓ ✓ Products for mass spec sample prep Protein ExtractionCell 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 Abundant Protein DepletionKits for albumin, top 2, or top 12 abundant protein depletion to aid in removing the most abundant proteins so that detection of lower abundant proteins is enhanced prior to digestion Protein EnrichmentEnrichment techniques include immunoprecipitation, enrichment by PTM (such as phosphoprotein enrichment), or active site labeling and enrichment using activity-based probes Peptide Quantitation AssaysThe colorimetric and fluorescent peptide quantitation assays help to normalize the peptide concentration of samples during each processing step following protein digestion Protein Digestion Using Mass Spec ProteasesProteases to break the proteins of interest into smaller fragments, or peptides for mass spec analysis Peptide Enrichment, Fractionation, and Clean-UpSuccessful analysis of low-abundant proteins and identification of post-translationally modified peptides requires multiple steps: enrichment, fractionation and/or clean up Protein Clean-UpA variety of specialty devices and resins that simply and efficiently desalt, buffer exchange and remove detergents from samples Protein Quantitation AssaysThe BCA protein assay is recommended prior to protein digestion to help normalize the protein concentration of samples during each processing step Related products for mass spec sample prep 24600,24615,24594 Resources Mass Spectrometry Reagents Support CenterPierce Protein MethodsProtein Biology Application NotesProtein Biology Selection GuidesOverview of Mass SpectrometryWorkflow for Protein Mass SpectrometrySample Preparation for Mass Spectrometry |
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