TMTpro™ 16plex Label Reagent Set, 1 x 5 mg, 1 x 5 mg (per tag) - FAQs

View additional product information for TMTpro™ 16plex Label Reagent Set - FAQs (A58334, A44520, A44521, A44522)

19 product FAQs found

What is the shelf life for the TMTpro 16plex Label Reagent Set?

The TMTpro 16plex Label Reagent Set is guaranteed for one year from the date of receipt in original, unopened packaging. This information can also be found in the Certificate of Analysis.

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Can the TMT and TMTpro reagents be mixed?

No, the TMT and TMTpro reagents cannot be mixed because the reporter ions are in the same region.

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Can TMTpro label reagents dissolved in anhydrous acetonitrile be dried down and stored for later use?

Yes, unused TMTpro label reagent that has been dissolved in anhydrous acetonitrile can be aliquoted into tubes, dried under vacuum at 25-30 degrees C, purged with nitrogen, and stored in a sealed bag with desiccant at -20 degrees C.

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What size formats is the TMTpro 16plex Label Reagent Set available in?

The TMTpro 16plex Label Reagent Set can be purchased in three different size formats:
1 x 0.5 mg (per tag), Cat No. A44521; sufficient for one 16plex isobaric experiment
6 x 0.5 mg (per tag), Cat No. A44522; sufficient for six 16plex isobaric experiments
1 x 5 mg (per tag), Cat No. A44520; sufficient for ten 16pex isobaric experiments

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If I already have TMTpro 16-plex tags, what do I need to order to get a total of 18-plex TMTpro tags?

Depending upon the number of vials and amount of each tag in the vial for the TMTpro 16-plex tags you already have, you will have to purchase the appropriate TMTpro-134C and TMTpro-135N Label Reagents.
- If you already have TMTpro 16plex Label Reagent Set, 1 x 5 mg (Cat. No. A44520), you will have to purchase TMTpro-134C and TMTpro-135N Label Reagents, 1 x 5 mg (Cat. No. A52046).
- If you already have TMTpro 16plex Label Reagent Set, 6 x 0.5 mg (Cat. No. A44522), you will have to purchase TMTpro-134C and TMTpro-135N Label Reagents, 6 x 0.5 mg (Cat. No. A52048).

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Do you offer stand-alone replacement tags for TMTpro 16plex Label Reagent Set (Cat. No. A44520)?

Unfortunately, we do not offer individual TMTpro tags as stand-alone products. TMTpro tags are only sold as part of a kit.

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At which step should I label my mass spec samples with TMT reagents?

TMT labeling can be performed after reduction, alkylation, digestion, or prior to cleanup, if the sample is buffered at appropriate pH (8-8.5) in a suitable buffer without primary amines (e.g., Tris, glycine). TMT labeling can also be performed after peptide clean-up. Labeling of peptides after clean-up enables measuring and normalizing of the peptide samples for equal mixing.

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Do I have to apply correction factors for analyzing data for TMT/TMTpro-labeled samples?

Applying correction factors is required for more accurate relative protein quantitation as the isotopic impurity is variable among the tags, ranging from 5-10%.

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How do I bridge the data from different sample sets for TMT-based quantitation?

This is most often done with the utilization of a "pool channel" for each multiplex set. This reference channel typically contains an equimolar mixture of all samples which is labeled with one of the TMT tags that is shared among the sets. The pool channel can be used to then normalize relative protein abundance measurements across multiplexed sets.

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In the EasyPep sample preparation workflow, it is mentioned that TMT/TMTpro reagent can be added to peptides before or after the clean-up step. Are there any differences between these two methods, regarding the labeling efficiency or relative quantitation?

The EasyPep kit chemistry is fully compatible with TMT/TMTpro reagents. Most commonly, TMT/TMTpro reagents are used to label peptides immediately after digestion and before peptide cleanup. This enables combining samples for a single cleanup step which reduces variability for more reproducible quantitative measurements. Another option is to label samples after peptide cleanup. This approach provides efficient labeling of the samples but may require the use of a peptide quantification assay to ensure that samples are equally mixed before LC-MS analysis. Both the workflows should provide high labelling efficiencies using our recommended TMT/TMTpro reagent to sample ratios.

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What type of mass spectrometer can I use for Tandem Mass Tag (TMT) analysis?

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.

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What is the main purpose of Tandem Mass Tag (TMT) reagents?

Tandem Mass Tag technology is used to individually label different protein samples so they can be combined into a single sample for LC-MS analysis. The major advantages of this workflow are higher sample throughput, less instrument analysis time, high precision of peptide quantitation among replicates, and fewer missing quantified proteins among different samples.

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Which downstream applications are compatible with the EasyPep Mini MS Sample Prep Kit?

The EasyPep Mini MS Sample Prep kit is compatible with TMT labeling, peptide fractionation, and phosphopeptide enrichment.

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Is TMT quantitation relative or absolute?

TMT quantitation is relative between samples unless a heavy peptide standard is included for comparison.

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What is the main purpose of TMT (Tandem Mass Tag) reagents?

TMT technology is used to perform multiplex quantitation of proteins extracted from cells and tissues. Rather than quantify single proteins (for example with an antibody), here, large numbers of proteins can be quantified in a single run.

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What is the Pierce TMT11plex Yeast Digest Standard and what is its main purpose?

The Pierce TMT11plex Yeast Digest Standard is a lyophilized yeast peptide mixture of four congenic strains labeled with TMT11plex reagents. It serves as a standard for quality assessment of LC and MS data for samples labeled with Tandem Mass Tag (TMT) reagents.

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When should I use quadrupole versus ion trap isolation?

Quadrupole isolation is faster because ions must travel through the quadrupole analyzer regardless of the analysis type chosen. As a result, there is little to no time penalty when using the quadrupole for MS2 isolation. In such a case, quadrupole transmission efficiency will depend on isolation width, m/z, and the cleanliness of the analyzer. Ion trap isolation is best suited for MS3 experiments, including those performed using multinotch isolation for TMT analysis; these types of isolation events are not possible with the quadrupole analyzer.

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How effective is the salt removal using Pierce Peptide Desalting Spin Columns?

With Pierce Peptide Desalting Spin Columns, more than 90% of salt or labeling tags such as TMT tags can be removed.

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What is the reproducibility of Pierce Peptide Desalting Spin Columns from sample to sample?

With Pierce Peptide Desalting Spin Columns, the reproducibility coefficient of variation (CV) is ± 20%.

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