Shop All Mass Tagging Systems for Proteomics

TMTsixplex™ Isobaric Label Reagent Set, 2 x 0.8 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.

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).

L-Lysine-2HCl, 4,4,5,5-D4 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 (4,4,5,5-D4), 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-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.

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

The carbonyl-reactive Thermo Scientific™ aminoxyTMT™ (Tandem Mass Tag™) Label Reagents enable multiplexed characterization and quantitation of carbonyl-containing biomolecules (carbohydrates, steroids, oxidized proteins) by mass spectrometry (MS).

The aminoxyTMTsixplex Label Reagents share an identical structure with aminoxyTMTzero Reagent but contain different numbers of 13C and 15N isotopes in the mass reporter. The optimized labeling procedure can be completed in one hour. After tag labeling, samples are quenched and cleaned-up using HILIC solid-phase extraction before mass spectrometry analysis.

The six additional compounds of the aminoxyTMTsixplex™ Reagent Set have the same mass (i.e., isobaric) and chemical structure (carbonyl-reactive aminoxy group, spacer arm, and mass reporter). However, the specific distribution of 13C and 15N isotopes on either side of the HCD or ETD MS/MS fragmentation site in each reagent results in a unique reporter mass (126-131Da) in the low mass region. This set of reporter ions is used to measure the relative abundance of labeled molecules in a combined (multiplexed) MS sample representing six different treatment conditions. For glycobiology MS applications, the reagents enable quantitative profiling of glycan isoforms and discovery of glycan biomarkers; they provide improved ionization of labeled glycans for increased sensitivity and better retention of labeled glycans by reversed-phase liquid chromatography (RPLC).The aminoxyTMT reagents may be used to quantify a broad range of biologically important molecules including carbohydrates, steroids, or oxidized proteins.

Features:

Quantitative – enables relative quantitation of glycans or other carbonyl-containing biomolecules from multiple samples derived from cells, tissues or biological fluids
Stable – oxime bond and products formed by labeling reaction are stable and don't require an additional reduction step
Efficient – achieve labeling efficiency greater than 90% in one hour
Sensitive – 20-fold increase in signal-to-noise compared to unlabeled native glycan MS analysis
Multiplex – able to identify and characterize up to six samples concurrently
Optimized – procedure and reagents optimized for excellent labeling efficiency and recovery of glycans

Includes:
• Label reagent sets contain sufficient aminoxyTMT tags for one sixplex experiment

Requires:
• PNGase F (or PNGase A) glycosidase and Waters Oasis™ HLB columns

Applications:
• Multiplex up to 6 different samples concurrently in a single experiment
• Relative quantitation of glycans
• Study of structural diversity of protein glycosylation
• Study of glycosylation in cell signaling and regulation
• Study of cancer progression, biomarker discovery and analysis of biotherapeutics

Native glycans are difficult to study by mass spectrometry because of their poor ionization efficiency. Quantitation of glycans is particularly challenging due to the lack of standards for all naturally-occurring glycans and difficulties reproducibly quantifying multiple samples. The aminoxy group has better reactivity with carbonyls and better stability of the labeled product compared to hydrazides. Labeling with the aminoxyTMT reagents improves ionization of glycans, thus improving sensitivity, and enables relative quantitation of glycans for up to six samples concurrently.

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.

These products are subject to a limited use label license.

Related Products
aminoxyTMTzero™ Label Reagent, 6 x 0.2 mg

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

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.

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

The Thermo Scientific SILAC Protein Quantitation Kit - 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:
• 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, 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.

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Dialyzed Fetal Bovine Sera, US Origin
Pierce Trypsin Protease, MS Grade
DMEM for SILAC

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.

Neucode™ Lysine-202 (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-202 (13C2 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-202 may be used with 4,4,5,5-D4 L-Lysine for NeuCode duplex experiments and may be combined with light lysine and 13C6 15N2 L-Lysine-2HCl for SILAC three-plex experiments.

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

NeuCode is a trademark of WARF.

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.

DMEM for SILAC (Thermo Scientific™)

Thermo Scientific™ DMEM 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 DMEM 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—medium is sterile, endotoxin-free, and cell culture-compatible

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 medium 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.

Related products
NeuCode™ Lysine -080, 50 mg
Fetal Bovine Serum, dialyzed, US origin
RPMI 1640 Media for SILAC

TMT10plex™ Isobaric Label Reagent Set, 8 x 0.2 mg (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.

Related Products
TMT10plex™ Isobaric Mass Tag Labeling Kit
1M Triethylammonium bicarbonate (TEAB) for TMT experiments
50% Hydroxylamine for TMT experiments

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

Powdered DMEM Medium for SILAC (Thermo Scientific™)

Thermo Scientific Powdered DMEM Medium 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 DMEM Medium for SILAC:

Flexible—powdered medium deficient in L-leucine, L-lysine and L-arginine, allowing for more complete proteome coverage through multiple isotopic amino acid labeling
High quality—medium is 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 medium is used together with heavy and light amino acids for the differential isotopic labeling of cells for SILAC analysis. SILAC medium supplemented with heavy amino acids are capable of nearly 100% label incorporation into proteins of living cells.

SILAC Protein Quantitation Kit (LysC), DMEM:F-12 (Thermo Scientific™)

The Thermo Scientific SILAC Protein Quantitation Kit - DMEM:F12 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:F12:
• 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:F12 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.

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.

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DMEM:F-12 for SILAC