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