The Thermo Scientific Pierce TMT11plex Yeast Digest Standard is a lyophilized yeast peptide mixture of four congenic strains labeled with TMT11plex reagents to monitor LC-MS/MS system performance for Tandem Mass Tag (TMT) quantitation.
Pierce TMT11plex Yeast Digest Standard features include:
—pre-labeled with TMT11plex reagents in a ready-to-use lyophilized format
• Highly validated
—yeast strains verified for genotype, equimolar mixing of the TMT-labeled strains, and overall protein quantitation using TMT
—excellent QC assay tool for quality assessment of LC and MS instrument status
—designed to measure the quantitative precision of the TMT reporter ions for each of the 11 channels
Quantitative proteomic strategies using TMT reagents enable sample multiplexing and precise measurement of protein abundance. However, successful execution of this workflow includes multiple steps that may require optimization, including chromatography, mass spectrometry (MS), and data analysis. A critical parameter for success in this workflow is the ability to consistently detect the TMT fragment ions across the 11 channels of multiplexing. This standard has been specifically designed as a tool to monitor LC-MS/MS system performance for TMT quantitation and MS method optimization and validation.
The Pierce TMT11plex Yeast Digest Standard consists of four congenic strains of Saccharomyces cerevisiae
(a parental line, BY4741, and three lines lacking the non-essential proteins Met6, His4, or Ura2) labeled in triplicate (knock-out strains) or duplicate (parental strain) with TMT11-plex label reagents (see figure). The knock-out strains are used in combination with the parental strain to enable comparison of peptide and protein abundance in the deleted and parental strains to monitor system performance for quantitation.
In addition to routine quality control assessment, the standard can also be used as a method development tool to measure the accuracy, precision, and dynamic range of different MS approaches, including optimization of offline fractionation, chromatography, MS acquisition, or data analysis settings.
The multifunctional applications for this standard will allow more reproducible TMT quantitation from day-to-day and experiment-to-experiment.References:
1. Paulo JA, O'Connell JD, Gygi SP. A Triple Knockout (TKO) Proteomics Standard for Diagnosing Ion Interference in Isobaric Labeling Experiments. J Am Soc Mass Spectrom. 2016 10:1620-5.
2. Yang F, Shen Y, Camp DG 2nd, Smith RD. High-pH reversed-phase chromatography with fraction concatenation for 2D proteomic analysis. Expert Rev Proteomics. 2012 9(2):129-34.
3. Dayon L, Pasquarello C, Hoogland C, Sanchez JC, Scherl A. Combining low- and high-energy tandem mass spectra for optimized peptide quantification with isobaric tags. J Proteomics. 2010 73(4):769-77.
4. Diedrich JK, Pinto AF, Yates JR 3rd. Energy dependence of HCD on peptide fragmentation: stepped collisional energy finds the sweet spot. J Am Soc Mass Spectrom. 2013 24(11):1690-9.
5. Chiva C, Sabidó E. HCD-only fragmentation method balances peptide identification and quantitation of TMT-labeled samples in hybrid linear ion trap/orbitrap mass spectrometers. J Proteomics. 2014 96:263-70.Related products:Pierce HeLa Protein Digest StandardPierce Peptide Retention Time Calibration MixtureTMT10plex isobaric label reagent set plus TMT11-131C label reagent