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  5. Thermo Scientific™ AccelerOme™ TMTpro™ 16plex MS Sample Preparation Kits

Thermo Scientific™ AccelerOme™ TMTpro™ 16plex MS Sample Preparation Kits

Catalog No. PIA50951
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Quantity:
1 x 16 TMTpro Reactions
2 x 16 TMTpro Reactions
Sufficient For:
10–30 μg samples
30–100 μg samples
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Catalog No. Quantity Sufficient For
PIA50951 2 x 16 TMTpro Reactions 30–100 μg samples
PIA50949 2 x 16 TMTpro Reactions 10–30 μg samples
PIA50950 1 x 16 TMTpro Reactions 30–100 μg samples
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Catalog No. PIA50951 Supplier Thermo Scientific™ Supplier No. A50951
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Description

TMTpro™ 16plex MS Sample Preparation Kits provide efficient and robust mass spec sample prep with TMTpro peptide labeling for use with the AccelerOme Automated Sample Prep System.

The AccelerOme TMTpro 16plex MS Sample Preparation Kits integrate pre-formulated buffers, MS-grade trypsin/Lys-C enzyme mix, and peptide clean-up cartridges with TMTpro 16plex label reagents for reproducible sample preparation of TMT-labeled peptides. The TMTpro 16plex label reagents have been optimized for use with samples from 10 to 100 μg samples to achieve greater than 95% labeling efficiency.

The Thermo Scientific Accelerome TMTpro 16plex MS Sample Preparation Kits provide a complete solution for efficient and robust proteomics mass spectrometry (MS) sample preparation, including TMT peptide labeling, for use with the AccelerOme Automated Sample Prep System.

TMTpro 16plex MS Sample Preparation Kits features:
• Flexible—sample prep and TMT-labeling reagents verified on system using cells, plasma, proteins, and tissue samples for 10 to 100 μg of sample input (kit dependent)
• Multiplex—enables concurrent MS analysis of up to 16 samples
• Optimized — EasyPep kit chemistry combined with TMTpro 16plex labeling reagents specifically designed for use with the AccelerOme Automated Sample Prep System
• Convenient—complete set of ready-to-use proteomic sample prep, clean-up, and TMT-labeling reagents for easy loading onto the instrument
• Robust —reagents combined with AccelerOme ecosystem provide reproducible, high-quality TMT-labeled peptides with greater than 95% labeling efficiency

Kit components are single-use and conveniently provided in labeled trays for direct placement on the AccelerOme system. This kit provides a complete reagent solution for automated sample preparation of TMTpro 16plex labeled samples to produce high quality, MS-ready peptides.

The AccelerOme reagent portfolio is an integral part of the AccelerOme Automated system solution to provide robust proteomic mass spec sample preparation. The instrument hardware is built on a proven liquid handling format for reproducible sample processing and optimized on-line μSPE sample clean-up for excellent peptide recovery. TMT label reconstitution and sample labeling and pooling have been incorporated before sample clean-up. Peptide quantitation post-clean-up is achieved through accurate UV spectrophotometric analysis to determine peptide concentrations prior to LC-MS analysis. The complete system (kits + reagents + software) was designed to provide high quality peptides while improving productivity with an easy-to-use automated system. When combined with the industry-leading, high-resolution Orbitrap instruments and software, TMTpro 16plex reagents provide integrated total solutions for quantitative protein expression analysis.

These products are subject to a limited use label license.
 

Specifications

Kit Contents Sample prep module, TMT module
Product Type Reagent Kit
Product Line Thermo Scientific
Sufficient For 30–100 μg samples
Starting Material Protein Samples
Final Product Type TMT-labeled peptide sample preparation
Detection Method UV Absorption
Format Kit
Workflow Step Automated mass spec sample prep
Quantity 2 x 16 TMTpro Reactions

Frequently Asked Questions (FAQs)

Can I use the AccelerOme TMT11plex MS Sample Preparation Kits and the AccelerOme TMTpro 16plex MS Sample Preparation Kits for samples with protein amounts lower than 10 µg?

Unfortunately, the AccelerOme TMT11plex MS Sample Preparation Kits and the AccelerOme TMTpro 16plex MS Sample Preparation Kits have not been validated for use with sample lysates containing lower than 10 µg of initial protein.

With the AccelerOme TMT11plex MS Sample Preparation Kits and the AccelerOme TMTpro 16plex MS Sample Preparation Kits, can I use any other lysis buffer instead of the one in the kit?

We do not recommend using a different lysis buffer other than the one included in the kit. On the one hand, variation in the lysis buffer might affect the subsequent sample prep steps. On the other hand, volume of the different buffers included in the kit has been optimized for the correct performance of the Accelerome platform.

When using the AccelerOme TMT11plex MS Sample Preparation Kits and the AccelerOme TMTpro 16plex MS Sample Preparation Kits with the AccelerOme platform, can I stop the process at any time?

We do not recommend stopping the automatized process as the AccelerOme platform is programmed to perform the complete sample preparation in a single run while keeping sample stability and reproducibility as its highest.

When using the AccelerOme TMT11plex MS Sample Preparation Kits and the AccelerOme TMTpro 16plex MS Sample Preparation Kits, should I add a protease inhibitor cocktail to the lysis buffer? How about phosphatase inhibitors?

We do not recommend using protease inhibitor cocktails containing EDTA with the AccelerOme TMT11plex MS Sample Preparation Kits and the AccelerOme TMTpro 16plex MS Sample Preparation Kits as they will inhibit both Universal nuclease and protease Trypsin/Lys-C activities. Following cell lysis, reduction and alkylation, the Trypsin/Lys-C proteases mix is added for complete digestion. Therefore, if protease inhibitors containing EDTA are added, there will be incomplete proteolysis. On the other hand, we do recommend using a phosphatase inhibitor cocktail (e.g., Halt Phosphatase Inhibitor Cocktail) if a subsequent phosphopeptide enrichment analysis will be performed.

Can I run the samples obtained from the AccelerOme TMT11plex MS Sample Preparation Kits and the AccelerOme TMTpro 16plex MS Sample Preparation Kits in any other mass spectrometer besides Orbitrap Mass Spectrometers?

We recommend using Orbitrap Mass Spectrometers as the analysis of TMT reagents requires a system that is able to perform high-resolution MS/MS fragmentation. Orbitrap Mass Spectrometers include a specific TMT MS2 workflow which enables unique acquisition management resulting in confident results without compromising coverage.

How should the samples be prepared prior to use of AccelerOme TMT11plex MS Sample Preparation Kits and the AccelerOme TMTpro 16plex MS Sample Preparation Kits?

Samples need to be resuspended into Lysis buffer and treated differently depending on the sample type (https://assets.thermofisher.com/TFS-Assets/LSG/manuals/MAN0025760_AccelerOmeTMT11plexMSSamplePrepandLabelingKits_UG.pdf). After determining protein concentration, 10-30 µg or 30-100 µg of each protein sample needs to be added into the appropriate wells of the Sample Input Plate and the volume should be adjusted up to 50 µL with Lysis buffer. Posterior clean-up, sample digestion, and TMT-labeling steps are automated.

How does the labeling occur in the AccelerOme TMT11plex MS Sample Preparation Kits and the AccelerOme TMTpro 16plex MS Sample Preparation Kits?

TMT labeling occurs over primary amines groups present at the N-terminal of peptides as well as on lysine residues.
Are the AccelerOme TMTpro 16plex MS Sample Preparation Kits, Cat. Nos. A50949 and A50951 designed for 32 different samples?

The AccelerOme TMTpro 16plex MS Sample Preparation Kits, Cat. Nos. A50949 and A50951 allow simultaneous running of 2 independent sets of experiments with 16 different samples in each set. Cat. No. A50949 is for samples containing 10 to 30 µg protein and Cat. No. A50951 is for samples containing 30 to 100 µg protein.
What is the difference between the AccelerOme TMTpro 16plex MS Sample Preparation Kits, Cat. Nos. A50949, A50950, and A50951?

The AccelerOme TMTpro 16plex MS Sample Preparation Kits, Cat. Nos. A50949, A50950, and A50951 are designed to prepare sets of 16 samples to be subjected to MS quantification. AccelerOme TMTpro 16plex MS Sample Preparation Kit, Cat. No. A50949 is designed for samples containing 10-30 µg protein whereas AccelerOme TMTpro 16plex MS Sample Preparation Kits, Cat. Nos. A50950 and A50951 are designed for samples containing 30-100 µg protein. Cat. Nos. A50949 and A50951 contain enough reagent and labels to perform 2 independent sets of 16 sample experiments whereas Cat. No. A50950 allows running of one single experiment with 16 samples.