Shop All Mass Tagging Systems for Proteomics

TMT11-131C Label Reagent (Thermo Scientific™)

Amine-reactive Thermo Scientific TMT11-131C increases sample multiplexing from 10-plex to 11-plex, enabling even greater throughput for protein identification and quantitative analysis by tandem mass spectrometry (MS).

Features of TMT11-131C:
Multiplex—increases level of multiplexing from ten to eleven samples
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 MS/MS platforms, such as the Q Exactive, Orbitrap Elite, and Orbitrap Fusion Tribrid instruments with data analysis fully supported by Proteome Discoverer 2.1

The Tandem Mass Tag (TMT) reagents are specially designed to enable identification and quantitation of proteins in different samples using tandem mass spectrometry. The TMT11-131C tag structure is identical to the other TMT10pex tags, but the TMT11-131C mass reporter is labeled with only 13C isotopes. The addition of the TMT11-131C tag enables separation of the 131 mass reporter ion of the original TMT10plex reagents into N and C variants, creating an 11th channel for relative quantitation using high resolution Orbitrap MS instruments.

Advantages of the TMT label reagents include increased multiplex relative quantitation, increased sample throughput, and fewer missing quantitative values among samples. TMT10plex label reagents plus TMT11-131C is ideal for analysis of multiple protein samples from inhibitor dose response experiments, time course experiments or biological replicates.

TMT11-131C is available here as a standalone tag or as part of theTMT10plex Isobaric Label Reagent Set plus TMT11-131C (Cat. No. A34808). When combined with the industry-leading, high resolution Orbitrap instruments and software, TMT reagents provide integrated total solutions for quantitative protein expression analysis.

Related products
TMT10plex Isobaric Label Reagent Set, 8 x 0.2 mg
TMT10plex Isobaric Label Reagent Set plus TMT11-131C, 1 x 5 mg
TMT10plex Isobaric Label Reagent Set, 1 x 5 mg

For high resolution analysis of the TMT-labeled peptides, the recommended LC column for Nanospray Flex source is Acclaim PepMap 100 C18 LC Column (Cat. No. 164942 or 164939). For EASY-Spray source, the recommended LC column is EASY-Spray C18 LC Column (Cat. No. ES802 or ES803).

TMTsixplex™ Isobaric Label Reagent Set, 2 x 5 mg (Thermo Scientific™)

Amine-reactive Thermo Scientific TMT™ Isobaric Mass Tagging Kits and Reagents enable multiplex quantitation of proteins extracted from cells and tissues using tandem mass spectrometry.

Tandem Mass Tag™ 6-plex (TMTsixplex™) Reagents are sets of isobaric compounds (i.e., same mass and structure, also called isotopomers) that are NHS-activated for covalent, irreversible labeling of primary amines (–NH2) groups. Each isobaric reagent contains a different number of heavy isotopes in the mass reporter region, which results in a unique reporter mass during tandem MS/MS for sample identification and relative quantitation. The reagents label all peptides prepared from cell or tissue samples for analysis of up to six samples in a single MS analysis.

Features of this TMTsixplex Label Reagent Set:

Powerful – concurrent MS analysis of multiple samples increases sample throughput and enables relative quantitation of up to six different samples derived from cells, tissues or biological fluids
Consistent – identical reagent structure and performance among TMTzero™, TMTduplex™, TMTsixplex™ and TMT10plex™ reagents allows efficient transition from method development to multiplex quantitation, enabling biomarker discovery research across platforms and datasets
Robust – consistent chemistry allows efficient transition from method development to multiplex quantitation, enabling biomarker discovery research
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 MS/MS platforms, such as the Q Exactive, Orbitrap Elite™ and Orbitrap Fusion™ Tribrid™ instruments with data analysis fully supported by Proteome Discoverer™ 1.0 and above

Applications:
• Protein identification and quantitation from multiple samples of cells, tissue or biological fluids
• Protein expression profiling of normal vs. disease states or control vs. treated
• Measurement of up to six different samples concurrently in a single experiment
• Quantitative analysis of proteins for which no antibodies are available
• Identification and quantitation of membrane and post-translationally modified proteins
• Identification and quantitation of hundreds to thousands of proteins in a single experiment

Changes in protein expression and post-translational modifications are essential mechanisms of biological regulation and disease. Advancements in liquid chromatography (LC), mass spectrometry (MS) instrumentation and bioinformatics now enable researchers to identify thousands of proteins in a given sample with a high degree of confidence.  However, reproducible detection and quantitation of different proteins within samples using mass spectrometry is challenging due to variability in peptide separation, ionization and detection.

Tandem Mass Tag (TMT) Reagents are isobaric chemical tags that enable concurrent identification and quantitation of proteins in different samples using tandem mass spectrometry. The amine-reactive, NHS-ester-activated compounds covalently attach to the peptide amino terminus and free amino termini of lysine residues of peptides and proteins with high efficiency, thereby labeling all peptides in a given sample regardless of enzyme used for digestion. Since TMT reagents share an identical structure and mass (i.e., isotopomers), labeled peptides co-elute during LC separation and are co-isolated during MS/MS analysis, resulting in fewer missing peptide identifications among samples. During MS/MS analysis, each isobaric tag is also fragmented to produce a unique reporter ion mass that is used for sample identification and quantitation. Protein quantitation is accomplished by comparing the relative intensities of the six reporter ions in the MS/MS spectra.

The Tandem Mass Tag (TMT) Reagent family consists of TMTzero, TMTduplex, TMTsixplex and TMT10plex sets which are specially designed to enable a rapid and cost-effective transition from method development to high-throughput protein quantitation. The TMTzero tag allows testing and optimization of sample preparation, labeling, fractionation and MS fragmentation for peptide identification and reporter detection without using the more costly isotope-labeled compounds. The TMTsixplex reagent set allows sixplex protein profiling for multiple conditions, including time courses, dose responses, replicates or multiple sample comparisons. Each TMT tag is based on the same chemical structure, eliminating the need to modify labeling conditions or HPLC separation conditions between experiments.

TMT Reagents are provided as standalone sets or in optimized kit formats containing all necessary reagents and controls for maximum flexibility, convenience and reliability. Tandem Mass Tag Reagents combined with the industry-leading Thermo Scientific instruments and software provide integrated total system solutions for quantitative protein expression analysis.

These products are subject to a limited use label license. Cell Signaling Technologies, IQ Proteomics, and OmicScouts are authorized service providers for tandem mass tags.

More Product Data
Quantitative cancer stem cell phosphoprotein profiling

Related Products
TMTsixplex™ Isobaric Mass Tagging Kit

For high resolution analysis of the TMT-labeled peptides, the recommended LC column for Nanospray Flex source is Acclaim PepMap 100 C18 LC Column (Cat. No. 164942 or 164939). For EASY-Spray source, the recommended LC column is EASY-Spray C18 LC Column (Cat. No. ES802 or ES803).

TMT10plex Isobaric Label Reagent Set plus TMT11-131C Label Reagent (Thermo Scientific™)

Amine-reactive Thermo Scientific TMT10plex Isobaric Mass Tag Labeling Reagent Set plus TMT11-131C increases sample multiplexing from 10-plex to 11-plex, enabling even greater throughput for protein identification and quantitative analysis by tandem mass spectrometry (MS).

Features of this TMT10plex Isobaric Label Reagent Set plus TMT11-131C:
Multiplex—concurrent MS analysis of up to 11 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 MS/MS platforms, such as the Q Exactive, Orbitrap Elite, and Orbitrap Fusion Tribrid instruments with data analysis fully supported by Proteome Discoverer 2.1

The Tandem Mass Tag (TMT) reagents are specially designed to enable identification and quantitation of proteins in different samples using tandem mass spectrometry. The TMT11-131C tag structure is identical to the other TMT10pex tags, but the TMT11-131C mass reporter is labeled with only 13C isotopes. The addition of the TMT11-131C tag enables separation of the 131 mass reporter ion of the original TMT10plex reagents into N and C variants, creating an 11th channel for relative quantitation using high resolution Orbitrap MS instruments.

Advantages of the TMT label reagents include increased multiplex relative quantitation, increased sample throughput, and fewer missing quantitative values among samples. TMT10plex label reagents plus TMT11-131C is ideal for analysis of multiple protein samples from inhibitor dose response experiments, time course experiments, or biological replicates.

TMT11-131C is available as a standalone tag (Cat. No. A34807), or as part of this TMT10plex Isobaric Label Reagent Set plus TMT11-131C. When combined with the industry-leading, high resolution Orbitrap instruments and software, TMT reagents provide integrated total solutions for quantitative protein expression analysis.

These products are subject to a limited use label license. Cell Signaling Technologies, IQ Proteomics, and OmicScouts are authorized service providers for tandem mass tags.

Related products
TMT10plex Isobaric Label Reagent Set, 8 x 0.2 mg
TMT11-131C, 1 x 5 mg
TMT10plex Isobaric Label Reagent Set, 1 x 5 mg

For high resolution analysis of the TMT-labeled peptides, the recommended LC column for Nanospray Flex source is Acclaim PepMap 100 C18 LC Column (Cat. No. 164942 or 164939). For EASY-Spray source, the recommended LC column is EASY-Spray C18 LC Column (Cat. No. ES802 or ES803).

NeuCode™ Lysine-602 (Thermo Scientific™)

Thermo Scientific NeuCode amino acids augment the level of multiplexing achievable for the metabolic labeling of proteins for mass spectrometry analysis. NeuCode Lysine-602 (13C6 15N2 L-Lysine-2HCl) may also be used with traditional SILAC to improve flexibility of multiplexing options or to reduce complexity of analysis.

General features of NeuCode SILAC labeling:
• Labeling efficiency—100% label incorporation into proteins of living cells without toxicity
• Compatible—may be multiplexed with existing SILAC amino acids
• Time-saving—not necessary to label to 100% incorporation if only using heavy amino acids
• High-quality supplements—heavy amino acids with >98% isotope purity

Stable isotope labeling with amino acids in cell culture (SILAC) is a powerful method to identify and quantify relative differential changes in complex protein samples. NeuCode metabolic labeling is similar to SILAC, but differs in that the labeling only utilizes heavy amino acids. The increased multiplexing capability of NeuCode amino acids is possible through the use of mass defects from extra neutrons in the stable isotopes. These small mass differences may be resolved on high resolution mass spectrometers (Thermo Scientific™ Orbitrap™ Elite™, Orbitrap™ Fusion™ Tribrid™, and Orbitrap™ Fusion™ Lumos™ Tribrid™ mass spectrometers). Use of only heavy amino acids eliminates the need for 100% incorporation of amino acids used for SILAC (both heavy and light), and may be especially useful for studies with primary cells.

NeuCode amino acids are used together with specialized cell culture media that are deficient in essential amino acids. Heavy L-lysine is used for SILAC analysis of peptides that have been digested with trypsin or LysC. NeuCode Lysine-602 may be used with NeuCode Lysine-080 for duplex experiments and may be combined with light lysine and 4,4,5,5-D4 L-lysine for three-plex experiments.

Related products
L-Lysine-2HCl, 4,4,5,5-D4
NeuCode Lysine-080
Neucode Lysine-341

NeuCode is a trademark of WARF.

1-Step Heavy Protein IVT Kit (Thermo Scientific™)

Thermo Scientific 1-Step Heavy Protein in vitro Protein Expression Kits enable rapid cell-free expression of recombinant proteins containing stable isotope-labeled (i.e., heavy) amino acids.

Features of the 1-Step Heavy Protein IVT Kit:

Efficient—express heavy proteins with 90 to 95% stable isotope incorporation
Functional—uses the human translational machinery to express more biologically active proteins than other IVT systems
Flexible—express light proteins or heavy proteins containing 13C615N2 L-lysine and/or 13C615N4 L-arginine
Convenient—perform transcription and translation in a single step
High performance—greater yields compared to rabbit reticulocyte in vitro translation

The 1-Step Heavy Protein IVT Kit uses a unique HeLa cell lysate supplemented with heavy amino acids for in vitro translation (IVT) of proteins with 90 to 95% isotope incorporation efficiency in less than 8 hours. Heavy proteins expressed using this system can be used as mass spectrometry controls for sample prep loss, digestion efficiency determination or as quantification standards.

Applications:
• Identify ideal peptides for targeted quantitation
• Verify protein digestion efficiency
• Control for protein sample prep variability and affinity enrichment loss

Thermo Scientific 1-Step Heavy Protein IVT Kit contains all of the necessary components to express a heavy protein including HeLa cell lysate, proprietary accessory proteins, reaction mix, heavy amino acids, positive-control GFP DNA and the pT7CFE1-CGST-HA-His cloning vector. The benefits of in vitro expression of heavy proteins over traditional in vivo systems include expression of toxic or insoluble proteins, a more rapid protein synthesis and a more economical use of heavy amino acids compared to stable-isotope labeled cell lines. This small-scale expression method makes it easy to express numerous heavy proteins simultaneously or to express large quantities of a single heavy protein up to 100 µg/mL. Tandem affinity purification of heavy proteins is aided using an expression vector containing multiple affinity tags including GST, HA and 6xHis.

More Product Data
Cell-free protein expression system for generating stable isotope-labeled proteins

NeuCode™ Lysine-440 (Thermo Scientific™)

Thermo Scientific NeuCode amino acids augment the level of multiplexing achievable for the metabolic labeling of proteins for mass spectrometry analysis. NeuCode Lysine-440 (3,4,5,6-13C4, 5,5,6,6-D4 L-Lysine-2HCl) may also be used with traditional SILAC to improve flexibility of multiplexing options or to reduce complexity of analysis.

General features of NeuCode SILAC labeling:
• Labeling efficiency—100% label incorporation into proteins of living cells without toxicity
• Compatible—may be multiplexed with existing SILAC amino acids
• Time-saving—not necessary to label to 100% incorporation if only using heavy amino acids
• High-quality supplements—heavy amino acids with >98% isotope purity

Stable isotope labeling with amino acids in cell culture (SILAC) is a powerful method to identify and quantify relative differential changes in complex protein samples. NeuCode metabolic labeling is similar to SILAC, but differs in that the labeling only utilizes heavy amino acids. The increased multiplexing capability of NeuCode amino acids is possible through the use of mass defects from extra neutrons in the stable isotopes. These small mass differences may be resolved on high resolution mass spectrometers (Thermo Scientific™ Orbitrap™ Elite™, Orbitrap™ Fusion™ Tribrid™, and Orbitrap™ Fusion™ Lumos™ Tribrid™ mass spectrometers). Use of only heavy amino acids eliminates the need for 100% incorporation of amino acids used for SILAC (both heavy and light), and may be especially useful for studies with primary cells.

NeuCode amino acids are used together with specialized cell culture media that are deficient in essential amino acids. Heavy L-lysine is used for SILAC analysis of peptides that have been digested with trypsin or LysC. NeuCode Lysine-440 may be used with 13C6 15N2 L-Lysine-2HCl (NeuCode Lysine-602), NeuCode Lysine-521, and NeuCode Lysine-080 for NeuCode four-plex experiments and may be combined with light lysine and 4,4,5,5-D4 L-lysine for three-plex experiments.

Related products
L-Lysine-2HCl, 4,4,5,5-D4
13C6 15N2 L-Lysine-2HCl
NeuCode Lysine-080
Neucode Lysine-521

NeuCode is a trademark of WARF.

iodoTMTsixplex™ Isobaric Label Reagent Set, 1 x 0.2 mg (Thermo Scientific™)

Thermo Scientific iodoTMTsixplex Label Reagents enable concurrent identification and multiplexed quantitation of proteins in different samples using tandem mass spectrometry. The iodoTMT™ Reagents are sets of isobaric (mass and structure) isomers that are iodoacetyl-activated for covalent, irreversible labeling of sulfhydryl (—SH) groups. Similar to iodoacetamide, iodoTMT Reagents react specifically with reduced cysteines (Cys) in peptides and proteins. IodoTMT Reagents can be differentiated by mass spectrometry (MS), enabling quantitation of the relative abundance of cysteine modifications, such as S-nitrosylation, oxidation and disulfide bonds, in cultured cells grown or treated with different conditions.

Features of iodoTMTsixplex Label Reagents:

Specific—only reacts with sulfhydryl groups (reduced, unmodified cysteine residues)
Irreversible—labeled proteins and peptides are not susceptible to reducing agents
Flexible—options for duplex isotopic (MS) or sixplex isobaric (MS/MS) quantitation
Versatile—can be used to study cysteine modifications (e.g. di-sulfide bridges, S-nitrosylation)

IodoTMT Products replace our previously offered CysTMT™ Reagents, which utilized a dithiopyridine reactive group to reversibly label cysteine sulfhydryls. For information about customized reagent sets, please contact our Large Volume and Custom Sales Department.

These products are subject to a limited use label license.

More Product Data
Specific labeling, enrichment and quantitation of S-nitrosylated peptides using iodoTMT reagents

Related Products
iodoTMTsixplex™ Isobaric Mass Tag Labeling Kit
iodoTMTzero™ Label Reagent, 5 x 0.2 mg
Immobilized Anti-TMT™ Antibody Resin
TMT™ Elution Buffer

SILAC Protein Quantitation Kit (LysC), RPMI 1640 (Thermo Scientific™)

The Thermo Scientific SILAC Protein Quantitation Kit - RPMI 1640 is used for the specific analysis of protein expression by mass spectrometry using stable isotope labeling with amino acids in cell culture (SILAC).

Features of the SILAC Protein Quantitation Kit - RPMI 1640:
• Efficient—up to 100% label incorporation into proteins of living cells
• Reproducible—minimizes intra-experimental variability caused by differential sample preparation
• Flexible—media deficient in both L-lysine and L-arginine, allowing for more complete proteome coverage through dual amino acid isotope labeling
• Compatible—label proteins expressed in a wide variety of mammalian cell lines adapted to grow in RPMI 1640, including HeLa, 293T, COS7, U2OS, A549, A431, HepG2, NIH 3T3, Jurkat and others
• High-quality supplements—heavy amino acids with >99% isotope purity; dialyzed FBS tested to help ensure that it is sterile, endotoxin-free, and cell culture compatible

Applications of SILAC Protein Quantitation Kit:
• Quantitative analysis of relative changes in protein abundance from different cell treatments
• Quantitative analysis of proteins for which there are no antibodies available
• Protein expression profiling to study normal vs. disease cells
• Identification and quantification of hundreds to thousands of proteins in a single experiment

SILAC is a powerful method to identify and quantify relative differential changes in complex protein samples. This approach entails the in vivo metabolic incorporation of "heavy" 13C- or 15N-labeled amino acids into proteins followed by mass spectrometry (MS) for the accelerated and comprehensive identification, characterization, and quantitation of proteins.

Because peptides labeled with heavy and light amino acids are chemically identical, they co-elute during reverse-phase column pre-fractionation and therefore are detected simultaneously during MS analysis. The relative peak intensities of multiple isotopically distinct peptides from each protein are then used to determine the average change in protein abundance in the treated sample.

Each kit includes all necessary reagents to isotopically label cells with 13C6 L-lysine-2HCl, including media, heavy and light amino acids, and dialyzed serum. Heavy L-arginine-HCl derivatives (Cat. Nos. 88210, 89990) are available separately and can be combined with Pierce SILAC Protein Quantitation kits to enhance peptide isotope label coverage. When combined with Thermo Scientific Protein/Peptide Sample Enrichment Products, Pierce SILAC Protein Quantitation kits also enable MS analysis of low-abundance proteins such as cell surface proteins, organelle-specific proteins, and post-translational protein modifications such as phosphorylation or glycosylation.

Related products
Dialyzed Fetal Bovine Sera, US Origin
Pierce Trypsin Protease, MS Grade
RPMI 1640 Medium for SILAC

NeuCode™ Lysine-521 (Thermo Scientific™)

Thermo Scientific NeuCode amino acids augment the level of multiplexing achievable for the metabolic labeling of proteins for mass spectrometry analysis. NeuCode Lysine-521 (1,2,3,4,5-13C5, 6,6-D2,15N L-Lysine-2HCl) may also be used with traditional SILAC to improve flexibility of multiplexing options or to reduce complexity of analysis.

General features of NeuCode SILAC labeling:
• Labeling efficiency—100% label incorporation into proteins of living cells without toxicity
• Compatible—may be multiplexed with existing SILAC amino acids
• Time-saving—not necessary to label to 100% incorporation if only using heavy amino acids
• High-quality supplements—heavy amino acids with >98% isotope purity

Stable isotope labeling with amino acids in cell culture (SILAC) is a powerful method to identify and quantify relative differential changes in complex protein samples. NeuCode metabolic labeling is similar to SILAC, but differs in that the labeling only utilizes heavy amino acids. The increased multiplexing capability of NeuCode amino acids is possible through the use of mass defects from extra neutrons in the stable isotopes. These small mass differences may be resolved on high resolution mass spectrometers (Thermo Scientific™ Orbitrap™ Elite™, Orbitrap™ Fusion™ Tribrid™, and Orbitrap™ Fusion™ Lumos™ Tribrid™ mass spectrometers). Use of only heavy amino acids eliminates the need for 100% incorporation of amino acids used for SILAC (both heavy and light), and may be especially useful for studies with primary cells.

NeuCode amino acids are used together with specialized cell culture media that are deficient in essential amino acids. Heavy L-lysine is used for SILAC analysis of peptides that have been digested with trypsin or LysC. NeuCode Lysine-521 may be used with 13C6 15N2 L-Lysine-2HCl (NeuCode Lysine-602), NeuCode Lysine-440, and NeuCode Lysine-080 for NeuCode four-plex experiments and may be combined with light lysine and 4,4,5,5-D4 L-Lysine for three-plex experiments.

Related products
L-Lysine-2HCl, 4,4,5,5-D4
13C6 15N2 L-Lysine-2HCl
NeuCode Lysine-080
Neucode Lysine-440

NeuCode is a trademark of WARF.

NeuCode™ Lysine-642 (Thermo Scientific™)

Thermo Scientific NeuCode amino acids augment the level of multiplexing achievable for the metabolic labeling of proteins for mass spectrometry analysis. NeuCode Lysine-642 (13C6; 5,5,6,6-D4; 15N2 L-Lysine:2HCl) may also be used with traditional SILAC to improve flexibility of multiplexing options or to reduce complexity of analysis.

General features of NeuCode SILAC labeling:
Labeling efficiency—100% label incorporation into proteins of living cells without toxicity
Compatible—may be multiplexed with existing SILAC amino acids
Time-saving—not necessary to label to 100% incorporation if only using heavy amino acids
High-quality supplements—heavy amino acids with >98% isotope purity

Stable isotope labeling with amino acids in cell culture (SILAC) is a powerful method to identify and quantify relative differential changes in complex protein samples. NeuCode metabolic labeling is similar to SILAC, but differs in that the labeling only utilizes heavy amino acids. The increased multiplexing capability of NeuCode amino acids is possible through the use of mass defects from extra neutrons in the stable isotopes. These small mass differences may be resolved on high resolution mass spectrometers (Thermo Scientific Orbitrap Elite, Orbitrap Fusion Tribrid, and Orbitrap Fusion Lumos Tribrid mass spectrometers). Use of only heavy amino acids eliminates the need for 100% incorporation of amino acids used for SILAC (both heavy and light), and may be especially useful for studies with primary cells.

NeuCode amino acids are used together with specialized cell culture media that are deficient in essential amino acids. Heavy L-lysine is used for SILAC analysis of peptides that have been digested with trypsin or LysC.

Related products
L-Lysine-2HCl, 4,4,5,5-D4
13C6 15N2 L-Lysine-2HCl
NeuCode Lysine-080
Neucode Lysine-440

NeuCode is a trademark of WARF.

TMT™ Elution Buffer (Thermo Scientific™)

Thermo Scientific TMT Elution Buffer is for use with Thermo Scientific Immobilized Anti-TMT Resin to enrich samples for peptides that have been chemically labeled with Tandem Mass Tag™ (TMT™) Reagents.

Immobilized Anti-TMT Resin and TMT Elution Buffer are effective for reducing sample complexity, improving dynamic range and studying cysteine modifications such as S-nitrosylation, oxidation and disulfide bonds of protein samples labeled with iodoTMT Reagents. This approach of selective thiol labeling and affinity enrichment is similar to isotope-coded affinity tags (ICAT™ Method) but allows for higher multiplex quantitation.

The Immobilized Anti-TMT Resin uses the highly specific Anti-TMT Antibody to bind peptides labeled with TMT Reagents. The antibody is specific for the mass reporter region of the TMT Reagents which allows for enrichment of TMT-labeled peptides regardless of peptide labeling chemistry (e.g., amine-reactive, sulfhydryl-reactive). The TMT Elution Buffer is a volatile neutral buffer which competitively elutes captured TMT-labeled peptides. When used in combination with the Immobilized Anti-TMT Resin, the TMT Elution Buffer provides higher specific enrichment of TMT-labeled peptides compared to acidic or basic elution buffers.

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Immobilized Anti-TMT™ Antibody Resin
iodoTMTsixplex™ Label Reagent Set, 1 x 0.2 mg
iodoTMTsixplex™ Isobaric Mass Tag Labeling Kit

Powdered RPMI Media for SILAC (Thermo Scientific™)

Thermo Scientific Powdered RPMI Media for SILAC is optimized for use with stable isotope labeling with amino acids in cell culture (SILAC) to analyze protein expression by mass spectrometry (MS).

Features of Powdered RPMI Media for SILAC:

Flexible—powdered media deficient in L-leucine, L-lysine and L-arginine, allowing for more complete proteome coverage through multiple isotopic amino acid labeling
High quality—media are sterile, endotoxin-free and cell culture-compatible

Stable isotope labeling with amino acids in cell culture (SILAC) is a powerful method to identify and quantify relative differential changes in complex protein samples. This approach entails the in vivo metabolic incorporation of "heavy" 13C- or 15N-labeled amino acids into proteins followed by mass spectrometry (MS) for the accelerated and comprehensive identification, characterization and quantitation of proteins.

This SILAC media is used together with heavy and light amino acids for the differential isotopic labeling of cells for SILAC analysis. SILAC media supplemented with heavy amino acids are capable of nearly 100% label incorporation into proteins of living cells.

TMTsixplex™ Isobaric Mass Tagging Kit (Thermo Scientific™)

Amine-reactive Thermo Scientific TMT™ Isobaric Mass Tagging Kits and Reagents enable multiplex quantitation of proteins extracted from cells and tissues using tandem mass spectrometry.

Tandem Mass Tag™ 6-plex (TMTsixplex™) Reagents are sets of isobaric compounds (i.e., same mass and structure, also called isotopomers) that are NHS-activated for covalent, irreversible labeling of primary amines (–NH2) groups. Each isobaric reagent contains a different number of heavy isotopes in the mass reporter region, which results in a unique reporter mass during tandem MS/MS for sample identification and relative quantitation. The reagents label all peptides prepared from cell or tissue samples for analysis of up to six samples in a single MS analysis.

Features of the TMTsixplex Isobaric Mass Tagging Kit:

Powerful – concurrent MS analysis of multiple samples increases sample throughput and enables relative quantitation of up to six different samples derived from cells, tissues or biological fluids
Consistent – identical reagent structure and performance among TMTzero™, TMTduplex™, TMTsixplex™ and TMT10plex™ reagents allows efficient transition from method development to multiplex quantitation, enabling biomarker discovery research across platforms and datasets
Robust – consistent chemistry allows efficient transition from method development to multiplex quantitation, enabling biomarker discovery research
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 MS/MS platforms, such as the Q Exactive, Orbitrap Elite™ and Orbitrap Fusion™ Tribrid™ instruments with data analysis fully supported by Proteome Discoverer™ 1.0 and above

Applications:
• Protein identification and quantitation from multiple samples of cells, tissue or biological fluids
• Protein expression profiling of normal vs. disease states or control vs. treated
• Measurement of up to six different samples concurrently in a single experiment
• Quantitative analysis of proteins for which no antibodies are available
• Identification and quantitation of membrane and post-translationally modified proteins
• Identification and quantitation of hundreds to thousands of proteins in a single experiment

Changes in protein expression and post-translational modifications are essential mechanisms of biological regulation and disease. Advancements in liquid chromatography (LC), mass spectrometry (MS) instrumentation and bioinformatics now enable researchers to identify thousands of proteins in a given sample with a high degree of confidence.  However, reproducible detection and quantitation of different proteins within samples using mass spectrometry is challenging due to variability in peptide separation, ionization and detection.

Tandem Mass Tag (TMT) Reagents are isobaric chemical tags that enable concurrent identification and quantitation of proteins in different samples using tandem mass spectrometry. The amine-reactive, NHS-ester-activated compounds covalently attach to the peptide amino terminus and free amino termini of lysine residues of peptides and proteins with high efficiency, thereby labeling all peptides in a given sample regardless of enzyme used for digestion. Since TMT reagents share an identical structure and mass (i.e., isotopomers), labeled peptides co-elute during LC separation and are co-isolated during MS/MS analysis, resulting in fewer missing peptide identifications among samples. During MS/MS analysis, each isobaric tag is also fragmented to produce a unique reporter ion mass that is used for sample identification and quantitation. Protein quantitation is accomplished by comparing the relative intensities of the six reporter ions in the MS/MS spectra.

The Tandem Mass Tag (TMT) Reagent family consists of TMTzero, TMTduplex, TMTsixplex and TMT10plex sets which are specially designed to enable a rapid and cost-effective transition from method development to high-throughput protein quantitation. The TMTzero tag allows testing and optimization of sample preparation, labeling, fractionation and MS fragmentation for peptide identification and reporter detection without using the more costly isotope-labeled compounds. The TMTsixplex reagent set allows sixplex protein profiling for multiple conditions, including time courses, dose responses, replicates or multiple sample comparisons. Each TMT tag is based on the same chemical structure, eliminating the need to modify labeling conditions or HPLC separation conditions between experiments.

TMT Reagents are provided as standalone sets or in optimized kit formats containing all necessary reagents and controls for maximum flexibility, convenience and reliability. Tandem Mass Tag Reagents combined with the industry-leading Thermo Scientific instruments and software provide integrated total system solutions for quantitative protein expression analysis.

More Product Data
Quantitative cancer stem cell phosphoprotein profiling

Related Products
TMTsixplex™ Label Reagent Set, 1 x 0.8 mg

SILAC Protein Quantitation Kit (Trypsin), DMEM (Thermo Scientific™)

The Thermo Scientific SILAC Protein Quantitation Kit (Trypsin), DMEM, is used for the specific analysis of protein expression by mass spectrometry using stable isotope labeling with amino acids in cell culture (SILAC).

Features of the SILAC Protein Quantitation Kit, DMEM, include:
• Efficient—up to 100% label incorporation into proteins of living cells
• Reproducible—minimizes intra-experimental variability caused by differential sample preparation
• Flexible—media deficient in both L-lysine and L-arginine, allowing for more complete proteome coverage through dual amino acid isotope labeling
• Compatible—label proteins expressed in a wide variety of mammalian cell lines adapted to grow in DMEM medium, including HeLa, 293T, COS7, U2OS, A549, A431, HepG2, NIH 3T3, Jurkat, and others
• High-quality supplements—heavy amino acids with >99% isotope purity; dialyzed FBS tested to help ensure that it is sterile, endotoxin-free, and cell culture compatible

Applications of SILAC Protein Quantitation Kit:
• Quantitative analysis of relative changes in protein abundance from different cell treatments
• Quantitative analysis of proteins for which there are no antibodies available
• Protein expression profiling to study normal vs disease cells
• Identification and quantification of hundreds to thousands of proteins in a single experiment

SILAC is a powerful method to identify and quantify relative differential changes in complex protein samples. This approach entails the in vivo metabolic incorporation of "heavy" 13C- or 15N-labeled amino acids into proteins followed by mass spectrometry (MS) for the accelerated and comprehensive identification, characterization, and quantitation of proteins.

Because peptides labeled with heavy and light amino acids are chemically identical, they co-elute during reverse-phase column pre-fractionation and therefore are detected simultaneously during MS analysis. The relative peak intensities of multiple isotopically distinct peptides from each protein are then used to determine the average change in protein abundance in the treated sample.

Trypsin SILAC kits are supplied with 13C6 15N L-lysine-2HCl and 13C6 15N4 L-arginine-HCl to label and quantify lysine-containing and arginine-containing peptides from trypsin protein digests. Each kit also includes 2 x 500 mL of medium and dialyzed serum. When combined with Thermo Scientific protein/peptide sample enrichment products, SILAC Protein Quantitation kits also enable MS analysis of low-abundance proteins such as cell surface proteins, organelle-specific proteins, and post-translational protein modifications such as phosphorylation or glycosylation.

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SILAC Protein Quantitation Kit (LysC), DMEM
SILAC Protein Quantitation Kit (LysC), DMEM:F-12
SILAC Protein Quantitation Kit (LysC), RPMI-1640

Ham's F12 Media for SILAC (Thermo Scientific™)

Thermo Scientific Ham's F12 Media for SILAC is optimized for use with stable isotope labeling with amino acids in cell culture (SILAC) to analyze protein expression by mass spectrometry (MS).

Features of Ham's F12 for SILAC:

Flexible—liquid media deficient in both L-lysine and L-arginine, allowing for more complete proteome coverage through multiple isotopic amino acid labeling
High quality—media are sterile, endotoxin-free and cell culture-compatible

Stable isotope labeling with amino acids in cell culture (SILAC) is a powerful method to identify and quantify relative differential changes in complex protein samples. This approach entails the in vivo metabolic incorporation of "heavy" 13C- or 15N-labeled amino acids into proteins followed by mass spectrometry (MS) for the accelerated and comprehensive identification, characterization and quantitation of proteins.

This SILAC media is used together with heavy and light amino acids for the differential isotopic labeling of cells for SILAC analysis. SILAC media supplemented with heavy amino acids are capable of nearly 100% label incorporation into proteins of living cells.