Thermo Scientific™ TMTpro™ 16plex Label Reagent Set

Catalog No.	Quantity
FERA58334	6 x 50 μg, 96 reactions
PIA44520	1 x 5 mg (per tag)
PIA44521	1 x 0.5 mg (per tag)
PIA44522	6 x 0.5 mg (per tag)

Catalog No. FERA58334 Supplier Thermo Scientific™ Supplier No. A58334

Description

The Thermo Scientific TMTpro 16plex Label Reagent Set enables multiplexing of up to 16 samples for multiplexed protein identification and quantitation using tandem mass spectrometry (MS/MS).

TMTpro label reagents are the next generation of tandem mass tags, designed to increase multiplexing without compromising protein identification and quantitation. The structures of the TMTpro and TMT tags are similar in being isobaric and amine reactive, but the TMTpro tag has a longer spacer region and isobutyl proline mass reporter. After fragmentation, each TMTpro tag generates a unique reporter mass (i.e., TMTpro 126-134Da) in the low-mass region of the high-resolution MS/MS spectrum used for multiplexed relative quantitation of protein expression.

The advantages of TMTpro tags include increased multiplexed relative quantitation, increased sample throughput, and fewer missing quantitative values among samples. With these capabilities, the TMTpro 16plex label reagents are suited for applications requiring higher multiplexing and fewer missing values, such as inhibitor dose response experiments, time course experiments, thermal shift assays, or single cell analysis. When combined with the industry leading, high-resolution Orbitrap instruments and software, TMTpro reagents provide integrated total solutions for quantitative protein expression analysis.

Features of TMTpro 16plex Label Reagent Sets include:
• Multiplex—concurrent MS analysis of up to 16 samples derived from cells, tissues, or biological fluids
• Robust—increased multiplex capability results in fewer missing quantitative values among samples and higher confidence among replicates
• Efficient—amine-reactive, NHS ester-activated reagents ensure efficient labeling of all peptides regardless of protein sequence or proteolytic enzyme specificity
• Compatible—optimized for use with high resolution Thermo Scientific Orbitrap MS/MS platforms, such as the Q Exactive and Orbitrap Fusion Tribrid instrument series, including the Orbitrap Eclipse Tribrid and Ascend and Orbitrap Exploris 480 mass spectrometers with data analysis fully supported by Proteome Discoverer 2.3

For high resolution analysis of TMTpro-labeled peptides, the recommended LC column for the Nanospray Flex source is the Acclaim PepMap 100 C18 LC Column (Cat. No. 164942 or 164939). For the EASY-Spray source, the recommended LC column is the EASY-Spray C18 LC Column (Cat. No. ES902 or ES003).

Specifications

• Formulation: Liquid in stabilized DMSO solution
• For Use With (Equipment): Mass Spectrometer
• Product Type: Isobaric Label Reagent Set
• Product Line: TMT
• Label or Dye: Tandem Mass Tag™ (TMT) Labels
• Starting Material: Cell Lysate
• Final Product Type: Peptide
• Detection Method: Mass Spectrometry
• Workflow Step: Protein Labeling
• Quantity: 6 x 50 μg, 96 reactions
• Form: Liquid

Frequently Asked Questions (FAQs)

How can I tell if the buffer in which my peptides are resuspended are compatible with TMT and TMTpro labels?

TMT and TMTpro labels are not compatible with buffers containing primary amines. After protein digestion, peptides need to be cleaned up and dried down. Prior to labeling, we do recommend resuspending the peptides in 10 µL of either 100 mM TEAB (pH 8.5) or 100 mM HEPES (pH 8.0).

Is the TMT stabilizing solution compatible with subsequent MS steps?

For further steps after TMT labelling, the stabilizing solution can be considered as a 33% DMSO solution. If subsequent excess TMT removal will be performed using EasyPep solid-phase extraction (SPE), the DMSO solution does not need to be diluted. On the other hand, the use of any reverse phase chromatography method (i.e., Cat. No. 84868) might need dilution of the sample to ensure that the DMSO concentration is tolerated by the resin. Alternatively, the peptide mix can be dried down using Speed Vac and then resuspended in the corresponding buffer for a particular column (Reverse Phase or Desalting).

With TMT 10plex, TMT 11plex, TMTpro 16plex, and TMTpro 18plex 96-well plates, how much protein can be used per TMT or TMTpro reaction?

TMT and TMTpro label reagents in 96-well plates are optimized to work with 1 to 10 µg of protein digest.

Can the sealing foil be partially removed with TMT 10plex, TMT 11plex, TMTpro 16plex, and TMTpro 18plex 96-well plates?

Yes, the sealing foil can be partially peeled to access individual sets of TMT or TMTpro labels. Alternatively, each of the wells can be perforated to transfer the TMT/TMTpro labels to a different tube, if needed.

How do I determine the correct orientation of the TMT 10plex, TMT 11plex, TMTpro 16plex, and TMTpro 18plex 96-well plates?

All the plates are barcoded in the long lower edge, helping to recognize correct plate orientation, plus the plate has a notch on the left corners.

Would the TMT and TMTpro labels be affected if the TMT 10plex, TMT 11plex, TMTpro 16plex, and TMTpro 18plex 96-well plates are going through several freeze-thaw cycles?

The TMT and TMTpro labels from TMT 10plex, TMT 11plex, TMTpro 16plex and TMTpro 18plex 96-well plates are stable after 9 freeze-thaw cycles, making it possible to use each of the experiment sets at different times.

With TMT 10plex, TMT 11plex, TMTpro 16plex, and TMTpro 18plex 96-well plates, can I run each of the experiment sets in different runs or should I use the plate all at once?

The entire plate can be run at one time or each experiment set can be run at a different time, using a high-throughput automation platform.

How many sets of experiments can I run with TMTpro 16plex Label Reagents ​in stabilized solution, ​96-well plate, 6 x 50 ug​ (Cat. No. A58334)?

TMTpro 16plex 96-well plate (Cat. No. A58334) contains 6 independent sets of experiments, each of them occupying two complete columns of the 96-well plate. Please see the user guide for further plate layout details.