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L-Lysine-2HCl, 13C6, 15N2 for SILAC (Thermo Scientific™)

Thermo Scientific Heavy and Light Amino Acids are used to specifically analyze protein expression by mass spectrometry using stable isotope labeling with amino acids in cell culture (SILAC) quantification kits. This is a lyophilized preparation of L-Lysine-2HCl (13C6, 15N2), sufficient for 1 SILAC experiment.

General features of heavy and light amino acids for SILAC labeling:

Efficient—100% label incorporation into proteins of living cells
Flexible—different isotopes of heavy and/or light amino acids for arginine, lysine, leucine and proline enable the quantitation of peptides derived from MS-grade proteases
Multiplex capabilities—several alternative isotopes of arginine and lysine are available that allow the analysis of multiple treatment conditions in each experiment
High-quality supplements—heavy amino acids with >99% 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. 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.

Thermo Scientific Heavy and Light Amino Acids for SILAC are used together with specialized cell culture media that are deficient in essential amino acids. Heavy and light L-lysine and L-arginine are the most common amino acids used for SILAC analysis of tryptic peptides. Up to three different experimental conditions can be readily analyzed with different isotopes of lysine and arginine. For lysine three-plex experiments, 4,4,5,5-D4 L-lysine and 13C6 15N2 L-lysine are used to generate peptides with 4- and 8-Da mass shifts, respectively, compared to peptides generated with light lysine. For arginine three-plex experiments, 13C6 L-arginine and 13C6 15N4 L-arginine are used to generate peptides with 6- and 10-Da mass shifts, respectively, compared to peptides generated with light arginine. L-leucine is another amino acid commonly used for SILAC labeling, because it is one of the most common amino acids found in protein sequences. Proline is a non-essential amino acid that is sometimes added to SILAC media to prevent the metabolic conversion of heavy arginine to heavy proline in mammalian cell lines with high arginine dehydrogenase activity.

Related Products
L-Arginine-HCl for SILAC
L-Proline for SILAC
L-Leucine for SILAC
L-Lysine-2HCl for SILAC

TMT10plex™ Isobaric Label Reagent Set, 3 x 0.8mg (Thermo Scientific™)

Amine-reactive Thermo Scientific TMT10plex™ Isobaric Mass Tag Labeling Reagents Sets enable multiplexed protein identification and quantitative analysis by tandem mass spectrometry (MS).

This Tandem Mass Tag™ (TMT™) 10-plex reagent set contains ten different isobaric compounds with the same mass and chemical structure (i.e., isotopomeric) composed of an amine-reactive NHS-ester group, a spacer arm and a mass reporter. The reagent set enables up to ten different peptide samples prepared from cells or tissues to be labeled in parallel and then combined for analysis. For each sample, a unique reporter mass (i.e., TMT10 126-131Da) in the low-mass region of the high-resolution MS/MS spectrum is used to measure relative protein expression levels during peptide fragmentation and tandem mass spectrometry.

Features of this TMT10plex Isobaric Label Reagent Set:

Powerful – concurrent MS analysis of multiple samples increases sample throughput and enables relative quantitation of up to ten 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 – 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™ 1.4

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
• Multiplex up to ten 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 quantification of hundreds to thousands of proteins in a single experiment

The Tandem Mass Tag (TMT) Reagents are specially designed to enable identification and quantitation of proteins in different samples using tandem mass spectrometry. The TMT10plex Label Reagents share an identical structure with TMTzero, TMTduplex, and TMTsixplex Reagents but contain different numbers and combinations of 13C and 15N isotopes in the mass reporter. The different isotopes result in a 10-plex set of tags that have mass differences in the reporter that can be detected using high resolution Orbitrap MS instruments.

Advantages of the TMT10plex Label Reagents include increased multiplex relative quantitation, increased sample throughput, and fewer missing quantitative values among samples. TMT10plex Label Reagents are ideal for analysis of multiple protein samples from inhibitor dose response experiments, time course experiments or biological replicates.

TMT Reagents are provided as standalone sets or in optimized kit formats containing all necessary reagents and controls for maximum flexibility, convenience and reliability. When combined with the industry-leading, high resolution Thermo Scientific Orbitrap instruments and software, TMT10plex 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 Mass Tag Labeling Kit
1M Triethylammonium bicarbonate (TEAB) for TMT experiments
50% Hydroxylamine for TMT experiments

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 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

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TMTsixplex™ Label Reagent Set, 1 x 0.8 mg

L-Arginine-HCl, 13C6 for SILAC (Thermo Scientific™)

Thermo Scientific Heavy and Light Amino Acids are used to specifically analyze protein expression by mass spectrometry using stable isotope labeling with amino acids in cell culture (SILAC) quantification kits. This is a lyophilized preparation of L-Arginine-HCl (13C6), sufficient for 1 SILAC experiment.

General features of heavy and light amino acids for SILAC labeling:

Efficient—100% label incorporation into proteins of living cells
Flexible—different isotopes of heavy and/or light amino acids for arginine, lysine, leucine and proline enable the quantitation of peptides derived from MS-grade proteases
Multiplex capabilities—several alternative isotopes of arginine and lysine are available that allow the analysis of multiple treatment conditions in each experiment
High-quality supplements—heavy amino acids with >99% 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. 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.

Thermo Scientific Heavy and Light Amino Acids for SILAC are used together with specialized cell culture media that are deficient in essential amino acids. Heavy and light L-lysine and L-arginine are the most common amino acids used for SILAC analysis of tryptic peptides. Up to three different experimental conditions can be readily analyzed with different isotopes of lysine and arginine. For lysine three-plex experiments, 4,4,5,5-D4 L-lysine and 13C6 15N2 L-lysine are used to generate peptides with 4- and 8-Da mass shifts, respectively, compared to peptides generated with light lysine. For arginine three-plex experiments, 13C6 L-arginine and 13C6 15N4 L-arginine are used to generate peptides with 6- and 10-Da mass shifts, respectively, compared to peptides generated with light arginine. L-leucine is another amino acid commonly used for SILAC labeling, because it is one of the most common amino acids found in protein sequences. Proline is a non-essential amino acid that is sometimes added to SILAC media to prevent the metabolic conversion of heavy arginine to heavy proline in mammalian cell lines with high arginine dehydrogenase activity.

Related Products
L-Arginine-HCl for SILAC
L-Proline for SILAC
L-Leucine for SILAC
L-Lysine-2HCl for SILAC

NeuCode™ Lysine-192 (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-192 (6-13C, D9, 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.

L-Proline for SILAC (Thermo Scientific™)

Thermo Scientific Heavy and Light Amino Acids are used to specifically analyze protein expression by mass spectrometry using stable isotope labeling with amino acids in cell culture (SILAC) quantification kits. This is a lyophilized preparation of L-Proline, sufficient for 10 SILAC experiments.

General features of heavy and light amino acids for SILAC labeling:

Efficient—100% label incorporation into proteins of living cells
Flexible—different isotopes of heavy and/or light amino acids for arginine, lysine, leucine and proline enable the quantitation of peptides derived from MS-grade proteases
Multiplex capabilities—several alternative isotopes of arginine and lysine are available that allow the analysis of multiple treatment conditions in each experiment
High-quality supplements—heavy amino acids with >99% 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. 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.

Thermo Scientific Heavy and Light Amino Acids for SILAC are used together with specialized cell culture media that are deficient in essential amino acids. Heavy and light L-lysine and L-arginine are the most common amino acids used for SILAC analysis of tryptic peptides. Up to three different experimental conditions can be readily analyzed with different isotopes of lysine and arginine. For lysine three-plex experiments, 4,4,5,5-D4 L-lysine and 13C6 15N2 L-lysine are used to generate peptides with 4- and 8-Da mass shifts, respectively, compared to peptides generated with light lysine. For arginine three-plex experiments, 13C6 L-arginine and 13C6 15N4 L-arginine are used to generate peptides with 6- and 10-Da mass shifts, respectively, compared to peptides generated with light arginine. L-leucine is another amino acid commonly used for SILAC labeling, because it is one of the most common amino acids found in protein sequences. Proline is a non-essential amino acid that is sometimes added to SILAC media to prevent the metabolic conversion of heavy arginine to heavy proline in mammalian cell lines with high arginine dehydrogenase activity.

Related Products
L-Arginine-HCl for SILAC
L-Proline for SILAC
L-Leucine for SILAC
L-Lysine-2HCl for SILAC

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.

TMTpro™ 16plex Label Reagent Set, 1 x 5 mg (Thermo Scientific™)

The Thermo Scientific TMTpro 16plex Label Reagent Set enables multiplexing of up to 16 samples for protein identification and quantitation using tandem mass spectrometry (MS/MS).
Features of this TMTpro 16plex Label Reagent Set 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 MS/MS platforms, such as the Q Exactive and Orbitrap Fusion Tribrid instrument series, including the Orbitrap Eclipse Tribrid and Orbitrap Exploris 480 mass spectrometers with data analysis fully supported by Proteome Discoverer 2.3

TMTpro label reagents are the next generation of tandem mass tags, designed to increase the level of sample multiplexing without compromising on protein identification and quantitation. The TMTpro tag structure is similar to the TMT tag structure in that the tags remain isobaric and amine reactive, but differs in the longer spacer region and isobutyl proline mass reporter region. After MS/MS 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 that is used for relative quantitation of protein expression levels.

The advantages of TMTpro tags include increased multiplex relative quantitation, increased sample throughput, and fewer missing quantitative values among samples. The TMTpro 16plex label reagents are suited for analyses of multiple protein samples, such as inhibitor dose response experiments, time course experiments, thermal shift assays, or biological replicates. When combined with the industry-leading, high-resolution Orbitrap instruments and software, TMTpro reagents provide integrated total solutions for quantitative protein expression analysis.

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. ES802 or ES803).These products are subject to a limited use label license.

NeuCode™ Lysine-390 (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-390 (3,4,5-13C3, D9 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.

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.

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

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).

TMTduplex™ Isobaric Mass Tagging Kit (Thermo Scientific™)

The Amine-reactive Thermo Scientific TMT™ Isobaric Mass Tagging Kit contains labeling reagents and associated reaction reagents that enable multiplex quantitation of proteins extracted from cells and tissues using tandem mass spectrometry.

Tandem Mass Tag™ 2-plex (TMTduplex™) 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 two samples in a single MS analysis.

Features of the TMTduplex Isobaric Mass Tagging Kit:

Powerful—concurrent MS analysis of two samples increases throughput and enables relative quantitation of two different treatment conditions
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 data sets
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 expression profiling of normal vs. disease states or control vs. treated
• Measurement of two 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

Tandem Mass Tag (TMT) Reagents are chemical tags that enable identification and quantitation of proteins in different samples using tandem mass spectrometry. The amine-reactive, NHS-ester-activatedcompounds 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 two reporter ions in the MS/MS spectra.

The TMTduplex isobaric reagent set allows protein profiling for two conditions using tandem MS analysis. 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 TMTduplex isotopic reagent set combines the TMTzero tag with one of the TMT6 tags for quantitative protein profiling of two conditions at the MS level similar to SILAC.

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.

Related Products
TMTduplex™ Isotopic Label Reagent Set, 5 x 0.8 mg
TMTduplex™ Isobaric Label Reagent Set, 5 x 0.8 mg
TMTzero™ Label Reagent, 5 x 0.8 mg

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

L-Lysine-2HCl, 13C6 for SILAC (Thermo Scientific™)

Thermo Scientific Heavy and Light Amino Acids are used to specifically analyze protein expression by mass spectrometry using stable isotope labeling with amino acids in cell culture (SILAC) quantification kits. This is a lyophilized preparation of L-Lysine-2HCl (13C6), sufficient for 10 SILAC experiments.

General features of heavy and light amino acids for SILAC labeling:

Efficient—100% label incorporation into proteins of living cells
Flexible—different isotopes of heavy and/or light amino acids for arginine, lysine, leucine and proline enable the quantitation of peptides derived from MS-grade proteases
Multiplex capabilities—several alternative isotopes of arginine and lysine are available that allow the analysis of multiple treatment conditions in each experiment
High-quality supplements—heavy amino acids with >99% 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. 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.

Thermo Scientific Heavy and Light Amino Acids for SILAC are used together with specialized cell culture media that are deficient in essential amino acids. Heavy and light L-lysine and L-arginine are the most common amino acids used for SILAC analysis of tryptic peptides. Up to three different experimental conditions can be readily analyzed with different isotopes of lysine and arginine. For lysine three-plex experiments, 4,4,5,5-D4 L-lysine and 13C6 15N2 L-lysine are used to generate peptides with 4- and 8-Da mass shifts, respectively, compared to peptides generated with light lysine. For arginine three-plex experiments, 13C6 L-arginine and 13C6 15N4 L-arginine are used to generate peptides with 6- and 10-Da mass shifts, respectively, compared to peptides generated with light arginine. L-leucine is another amino acid commonly used for SILAC labeling, because it is one of the most common amino acids found in protein sequences. Proline is a non-essential amino acid that is sometimes added to SILAC media to prevent the metabolic conversion of heavy arginine to heavy proline in mammalian cell lines with high arginine dehydrogenase activity.

Related Products
L-Arginine-HCl for SILAC
L-Proline for SILAC
L-Leucine for SILAC
L-Lysine-2HCl for SILAC

NeuCode™ Lysine-341 (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-341 (3,4,5-13C3, 5,5,6,6-D4, 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-341 may be used with13C6 15N2 L-Lysine 2HCl (NeuCode Lysine-602) and NeuCode Lysine-080 for NeuCode three-plex experiments and may be combined with light lysine and 4,4,5,5-D4 L-Lysine for SILAC three-plex experiments.

Related products
L-Lysine-2HCl, 4,4,5,5-D4
13C6 15N2 L-Lysine-2HCl
NeuCode Lysine-080
Neucode Lysine-202

NeuCode is a trademark of WARF.