AccelerOme™ TMT11plex MS Sample Preparation Kit, 30–100 μg, 1 x 11 TMT Reactions - FAQs

View additional product information for AccelerOme™ TMT11plex MS Sample Preparation Kits - FAQs (A50946, A50947, A50948)

9 product FAQs found

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.

Find additional tips, troubleshooting help, and resources within our Mass Spectrometry Support Center.

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.

Find additional tips, troubleshooting help, and resources within our Mass Spectrometry Support Center.

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.

Find additional tips, troubleshooting help, and resources within our Mass Spectrometry Support Center.

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.

Find additional tips, troubleshooting help, and resources within our Mass Spectrometry Support Center.

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.

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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.

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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.

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Are the AccelerOme TMT11plex MS Sample Preparation Kits, Cat. Nos. A50946 and A50948 designed for 33 different samples?

The AccelerOme TMT11plex MS Sample Preparation Kits, Cat. Nos. A50946 and A50948 allow simultaneous running of 3 independent sets of experiments with 11 different samples in each set. Cat. No. A50946 is for samples containing 10 to 30 µg protein and Cat. No. A50948 is for samples containing 30 to 100 µg protein.

Find additional tips, troubleshooting help, and resources within our Mass Spectrometry Support Center.

What is the difference between the AccelerOme TMT11plex MS Sample Preparation Kits, Cat. Nos. A50946, A50947, and A50948?

The AccelerOme TMT11plex MS Sample Preparation Kits, Cat. Nos. A50946, A50947, and A50948 are designed to prepare sets of 11 samples to be subjected to MS quantification. AccelerOme TMT11plex MS Sample Preparation Kit, Cat. No. A50946 is designed for samples containing 10-30 µg protein whereas AccelerOme TMT11plex MS Sample Preparation Kits, Cat. Nos. A50947and A50948 are designed for samples containing 30-100 µg protein. Cat. Nos. A50946 and A50948 contain enough reagent and labels to perform 3 independent sets of 11 sample experiments whereas Cat. No. A50947 allows running of one single experiment with 11 samples.

Find additional tips, troubleshooting help, and resources within our Mass Spectrometry Support Center.