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Description
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. The knock-out strains are used in combination with the parental strain to enable comparison of peptide and protein abundance for quantitation. Pierce TMT11plex Yeast Digest Standard:
• Convenient—ready-to-use lyophilized powder pre-labeled with TMT11plex reagents
• Quality—yeast strains verified for genotype, equimolar mixing of the TMT-labeled strains
• Quantitative—measure the quantitative precision of TMT reporter ions for 11 channels
• Diagnostic—excellent QC assay tool for protein quantitation using TMT on LC-MS/MS instruments
Quantitative proteomic strategies using TMT reagents enable multiplexing and precise measurement of protein abundance. Successful execution of TMT quantitation includes optimized chromatography, mass spectrometry (MS), and data analysis. A critical parameter for success in this workflow is the ability to consistently detect TMT fragment ions across 11 channels for multiplexing. Our TMT11plex Yeast Digest standard has been specifically designed to assess LC-MS/MS system performance for TMT quantitation, MS method optimization and system validation. For method development, TMT11plex Yeast Digest Standard can be used to measure the accuracy, precision, and dynamic range of different mass spectrometry approaches. These standards can help optimize offline fractionation, chromatography, mass spectrometry acquisition, and data analysis settings. Multifunctional applications using this standard reduces inter-experiment variability to provides reproducible TMT quantitation.
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 Standard (Cat. No. 88329)
Pierce Peptide Retention Time Calibration Mixture (Cat. No. 88321)
TMT10plex isobaric label reagent set plus TMT11-131C label reagent (Cat. No. A34808)
Specifications
Specifications
| For Use With (Equipment) | Mass Spectrometer |
| Product Type | Digest Standard |
| Product Line | Pierce |
| Shipping Condition | Wet Ice |
| Starting Material | Peptides |
| Final Product Type | MS Standard |
| Detection Method | Mass Spectrometry |
| Workflow Step | MS System Performance |
| Quantity | 5 x 20 μg |
| Form | Lyophilized Powder |
Frequently Asked Questions (FAQs)
Here are some recommendations:
- Clean and recalibrate the mass spectrometry instrument using one of our Pierce Calibration Solutions (https://www.thermofisher.com/search/browse/category/us/en/600654).
- Verify settings for liquid chromatography (LC) acquisition methods.
- Fractionate TMT-labeled samples using the Pierce High pH Reversed-Phase Peptide Fractionation Kit (Cat. No. 84868) to reduce sample complexity.
The Pierce TMT11plex Yeast Digest Standard (i.e., TKO standard) is a lyophilized yeast peptide mixture of four congenic strains labeled with TMT11plex reagents. It can be used to rapidly and accurately assess ion interference (co-isolation of multiple analytes of similar mass-to-charge) for TMT-based quantitative mass spectrometry proteomic analysis.
We recommend using our Pierce TMT11plex Yeast Digest Standard (Cat. Nos. A40938, A40939) for LC-MS method optimization and for assessing system suitability for TMT-labeled samples.
Here are our recommendations:
- Pierce Peptide Retention Time Calibration Mixture (Cat. No. 88321) for LC system and gradient optimization
- Pierce BSA Protein Digest, MS grade (Cat. No. 88341), Pierce 6 Protein Digest, equimolar, LC-MS grade (Cat. No. 88342), and Pierce HeLa Protein Digest Standard (Cat. Nos. 88328, 88329) for "bottom up" LC-MS method optimization and LC-MS suitability assessment
- Pierce LC-MS/MS System Suitability Standard (7 x 5 Mix) (Cat. No. A40010) for assessing the gradient, sensitivity, and dynamic range of the LC-MS system for discovery and targeted peptide quantitative workflows
- Pierce TMT11plex Yeast Digest Standard (Cat. Nos. A40938, A40939) for assessing system performance for TMT-based quantitative workflows
- Pierce Intact Protein Standard Mix (Cat. Nos. A33526, A33527) for "top down" LC-MS method optimization and LC-MS suitability assessment
We recommend using high-resolution Orbitrap Tribrid (e.g., Fusion, Lumos, Eclipse), Orbitrap Exploris (e.g., 240, 480), or Q Exactive (e.g,. Plus, HF, HFX) series of instruments.
For Research Use Only. Not for use in diagnostic procedures.
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