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NeuCode™ Lysine-080 (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-080 (3,3,4,4,5,5,6,6-D8 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™, Q Exactive™, 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-080 may be used with 4,4,5,5-D4 L-lysine or 13C6 15N2 L-lysine for duplex experiments and may be combined with light lysine for three-plex experiments.

Related products
L-Lysine-2HCl, 4,4,5,5-D4 for SILAC
L-Lysine-2HCl, 13C6, 15N2 for SILAC
Fetal Bovine Serum, dialyzed, US origin
DMEM Media for SILAC

NeuCode is a trademark of WARF.

Pierce™ Top 2 Abundant Protein Depletion Spin Columns (Thermo Scientific™)

Thermo Scientific Pierce Abundant Protein Depletion Spin Columns are optimized to decrease the abundant albumin and antibody components of human plasma samples in preparation for mass spectrometry or 1D and 2D gel electrophoresis.

Features of Top 2 Abundant Protein Depletion Spin Columns:

Optimized—resin in spin columns is scaled and optimized for treating 10 µL (600 µg) of human plasma samples for mass spectrometry and/or 1D and 2D electrophoresis
Efficient—kits remove >90% of IgG and >95% of albumin, plus other abundant proteins (Top 12)
Fast—process samples in 40 to 60 minutes (depending on resin)
Economical—cost effective spin columns are priced for single use
Reduce variability—One time use prevents abundant protein carryover and experimental variability
Two resin options—choose the system appropriate for your need: 2 or 12 protein depletion columns

The Pierce Top 2 Abundant Protein Depletion Spin Columns use highly specific immobilized anti-HSA and anti-IgG antibodies to remove human serum albumin (HSA) and all major subclasses of gamma globulin (IgG) from serum, plasma or spinal fluids. The resins are provided in an economical and convenient spin column specifically designed one step processing and for single use.

Analysis of human fluids is often complicated by the presence of high concentrations of albumin and IgG that can make up more than 70% of total serum protein. Removal of these proteins is often essential for the study of low-abundant proteins. The Pierce Top 2 Protein Depletion Columns can deplete greater than 99% of HSA and 90% of IgG. The Pierce Top 12 Protein Depletion Columns can deplete greater than 95% of the twelve most abundant proteins in serum. Samples are loaded in a pre-filled convenient disposable spin format and can be processed in 40 to 60 minutes. The depletion of highly abundant proteins enables the detection of low abundant proteins in samples and subsequent identification by mass spectrometry or 1D and 2D gel electrophoresis.

Related Products
Pierce™ Top 12 Abundant Protein Depletion Spin Columns

SureQuant™ AKT Isoform Panel (Relative Quantitation) (Thermo Scientific™)

The Thermo Scientific SureQuant AKT Isoform Panel (Relative Quantitation) provides a complete reagent and quantitation solution for the enrichment and analysis of AKT1, AKT2, and AKT3 isoforms using mass spectrometry (MS).

The SureQuant AKT Isoform Panel (Relative Quantitation) consists of two modules:
SureQuant AKT Isoform IP and MS Sample Preparation Module includes all of the reagents necessary to immuno-enrich AKT1, AKT2, and AKT3 proteins and perform in-solution MS sample preparation from sample lysis through peptide clean-up
SureQuant AKT Isoform Relative Quantitation Module includes a system suitability standard to monitor MS performance, AQUA Ultimate HeavyPeptide mixture for relative peptide quantitation, and methods for using Thermo Scientific Q Exactive and Fusion Tribrid instruments and Nanospray LC instrumentation

SureQuant AKT Isoform Panel features include:
Complete—includes all reagents for successful monitoring of MS system performance, sample preparation, and absolute quantitation of target proteins/peptides
Validated—antibodies, peptides, and control lysate rigorously tested for specificity and successful quantitation of each target peptide
Multiplex—able to quantitate isoforms and phosphorylation of AKT
Flexible—modular format allows for immuno-enrichment only, or in combination with relative or absolute quantitation

The AKT signaling pathway serves as a master regulatory pathway for cell growth and proliferation, and AKT plays a vital role in the signaling cascade. Aberrant activation of this pathway promotes survival of tumor cells in many types of cancer. This kit is ideal for monitoring AKT expression levels of the three isoforms and phosphorylation status in a single assay. The ability to simultaneously monitor and quantitate in a single sample is a powerful tool for assessment of normal and abnormal pathway signaling.

The SureQuant AKT Isoform IP and MS Sample Preparation Module enables highly effective antigen immunoprecipitation (IP) and co-immunoprecipitation (co-IP) for MS analysis. Cells are first lysed and incubated with an IP-MS-verified biotinylated antibody mixture. The antibody-antigen complex is then captured on MS-compatible streptavidin magnetic beads, where non-specific proteins are removed through washing with MS-compatible buffers. The target proteins are eluted using trypsin and are directly compatible with MS sample preparation. The MS sample preparation has been streamlined and can be completed in ~4 hours for same-day MS analysis.

The SureQuant AKT Isoform Relative Quantitation Module provides reagents for relative quantitation of six unique peptides from AKT1, AKT2, and AKT3 isoforms using liquid chromatography (LC) and MS. AQUA Ultimate HeavyPeptide mixture is provided for use as a spike-in internal standard for relative quantitation in the multiplexed sample. Standardized methods and data process templates are provided for streamlined data acquisition and analysis.

The Pierce LC-MS/MS System Suitability Standard (7 x 5 Mix) is included in this module to assess dynamic range of LC-MS/MS systems before running the calibration curves and samples.

Together, these modules provide a complete solution for sample preparation and analysis of the target proteins.

Related products:
EASY-Spray LC Column
SureQuant AKT Pathway (Phospho) Multiplex Panel (Absolute Quantitation)
Pierce Trypsin Protease, MS Grade
Pierce LC-MS/MS System Suitability Standard

Pierce™ Top 12 Abundant Protein Depletion Spin Columns (Thermo Scientific™)

Thermo Scientific Pierce Abundant Protein Depletion Spin Columns are optimized to decrease the abundant albumin and antibody components of human plasma samples in preparation for mass spectrometry or 1D and 2D gel electrophoresis.

Features of Top 12 Abundant Protein Depletion Spin Columns:

Optimized—resin in spin columns is scaled and optimized for treating 10 µL (600 µg) of human plasma samples for mass spectrometry and/or 1D and 2D electrophoresis
Efficient—kits remove >90% of IgG and >95% of albumin, plus other abundant proteins (Top 12)
Fast—process samples in 40 to 60 minutes (depending on resin)
Economical—cost effective spin columns are priced for single use
Reduce variability—One time use prevents abundant protein carryover and experimental variability
Two resin options—choose the system appropriate for your need: 2 or 12 protein depletion columns

The Pierce Top 12 Abundant Protein Depletion Spin Columns are designed to remove HSA, IgG and 10 other high-abundance proteins from human serum or plasma. The resins are provided in an economical and convenient spin column specifically designed one step processing and for single use.

Analysis of human fluids is often complicated by the presence of high concentrations of albumin and IgG that can make up more than 70% of total serum protein. Removal of these proteins is often essential for the study of low-abundant proteins. The Pierce Top 2 Protein Depletion Columns can deplete greater than 99% of HSA and 90% of IgG. The Pierce Top 12 Protein Depletion Columns can deplete greater than 95% of the twelve most abundant proteins in serum. Samples are loaded in a pre-filled convenient disposable spin format and can be processed in 40 to 60 minutes. The depletion of highly abundant proteins enables the detection of low abundant proteins in samples and subsequent identification by mass spectrometry or 1D and 2D gel electrophoresis.

Related Products
Pierce™ Top 2 Abundant Protein Depletion Spin Columns

SureQuant™ AKT Isoform IP and MS Sample Preparation Module (Thermo Scientific™)

The Thermo Scientific SureQuant AKT Isoform IP and MS Sample Preparation Module provides a complete reagent solution for the multiplexed enrichment and sample preparation of AKT1, AKT2, and AKT3 for mass spectrometry (MS).

SureQuant AKT Isoform IP and MS Sample Preparation Module features include:
Complete—includes all of the reagents necessary for sample lysis, immuno-enrichment, and MS sample preparation
Validated—antibodies validated for specificity and background
Optimized—procedure simplified and condensed to prepare samples after immunoprecipitation in four hours
Multiplex—able to immuno-enrich AKT1, AKT2, and AKT3 isoforms in a single reaction

The AKT signaling pathway serves as a master regulatory pathway for cell growth and proliferation, and AKT plays a vital role in the signaling cascade. Aberrant activation of this pathway promotes survival of tumor cells in many types of cancer. This module is ideal for immuno-enrichment of the AKT1, AKT2, and AKT3 isoforms for use with the SureQuant quantitation modules.

The SureQuant AKT Isoform IP and MS Sample Preparation Module enables highly effective antigen immunoprecipitation (IP) and co-immunoprecipitation (co-IP) for MS analysis. Cells are first lysed and incubated with an IP-MS-verified biotinylated antibody mixture. The antibody-antigen complex is then captured on MS-compatible streptavidin magnetic beads, where non-specific proteins are removed through washing with MS-compatible buffers. The target proteins are eluted using trypsin and are directly compatible with MS sample preparation. The MS sample preparation has been streamlined and can be completed in ~4 hours for same-day MS analysis.

This kit can be used with the either the SureQuant AKT Isoform Absolute Quantitation Module or the SureQuant AKT Isoform Relative Quantitation Module for protein targeted quantitation. When used together, these modules provide a complete solution for sample preparation and analysis of the target proteins.

Related products:
EASY-Spray LC Column
SureQuant AKT Pathway Multiplex Panel (Absolute Quantitation)
Pierce Trypsin Protease, MS Grade
Pierce LC-MS/MS System Suitability Standard

MSIA™ D.A.R.T.'S™ for the Versette and Novus i Platform (Thermo Scientific™)

Improve hybrid MS bio-analysis with Thermo Scientific™ MSIA™ D.A.R.T.'S™ for the Versette and Novus i Platform an exclusive Thermo Fisher Scientific workflow solution for proteins and peptides.

SureQuant™ AKT Isoform Relative Quantitation Module (Thermo Scientific™)

The Thermo Scientific SureQuant AKT Isoform Relative Quantitation Module provides a complete quantitation solution for the analysis of key AKT expression using mass spectrometry (MS).

SureQuant AKT Isoform Relative Quantitation Module features include:
Complete—contains validated AQUA HeavyPeptide mix, diluent, matrix, system suitability standard including optimized acquisition and analysis methods
Validated—peptides rigorously tested for successful quantitation of each target peptide
Multiplex—able to quantitate AKT1, AKT2, and AKT3 isoforms and phosphorylation of AKT

The AKT signaling pathway serves as a master regulatory pathway for cell growth and proliferation, and AKT plays a vital role in the signaling cascade. Aberrant activation of this pathway promotes survival of tumor cells in many types of cancer. This kit is ideal for monitoring AKT expression levels of the three isoforms and phosphorylation status in a single assay. The ability to simultaneously monitor and quantitate in a single sample is a powerful tool for assessment of normal and abnormal pathway signaling.

The SureQuant AKT Isoform Relative Quantitation Module provides reagents for relative quantitation of six unique peptides from AKT using liquid chromatography (LC) and MS. AQUA Ultimate HeavyPeptide mixture is provided for use as a spike-in internal standard for relative quantitation in the multiplexed sample. Standardized methods and data process templates are provided for streamlined data acquisition and analysis.

The Pierce LC-MS/MS System Suitability Standard (7 x 5 Mix) is included in this module to assess dynamic range of LC-MS/MS systems before running the calibration curves and samples.

This kit can be used with SureQuant AKT Isoform IP and MS Sample Preparation Module for enriched multiplex sample preparation. When used together, these modules provide a complete solution for sample preparation and analysis of the target proteins.

Related products:
EASY-Spray LC Column
SureQuant AKT Pathway (Phospho) Multiplex Panel (Absolute Quantitation)
Pierce Trypsin Protease, MS Grade
Pierce LC-MS/MS System Suitability Standard

DSSO (disuccinimidyl sulfoxide) (Thermo Scientific™)

DSSO (disuccinimidyl sulfoxide) is an MS-cleavable and membrane-permeable crosslinker that contains an amine-reactive N-hydroxysuccinimide (NHS) ester at each end of a 7-carbon spacer arm. It facilitates analysis of protein structure and complex interactions using mass spectrometry. DSSO has similar reactivity to DSS and BS3, but contains a linker that can be cleaved in the gas phase during tandem MS (MS/MS) using collision-induced dissociation (CID). The ability to cleave cross-linked peptides during MS/MS enables MS3 acquisition methods, which facilitate peptide sequencing using traditional database search engines. The MS cleavage of DSSO also generates diagnostic ion doublets during MS2, which enables identification of cross-linked peptides from dead-end modifications and searching using novel database search engines such as XlinkX.

Features of DSSO:
• Reactive groups: NHS ester (both ends)
• Reactive towards: amino groups (primary amines)
• Amine-reactive NHS ester reacts rapidly with any primary amine-containing molecule
• Membrane permeable, allowing for intracellular crosslinking
• High-purity, crystalline reagent can be used to create high-purity conjugates
• MS-cleavable
• Water-insoluble (dissolve first in DMF or DMSO)

Lot specifications:
• Purity: > 90% by quantitative NMR (the highest standard in crosslinking purity)

Chemical crosslinking in combination with mass spectrometry is a powerful method to determine protein-protein interactions. This method has been applied to recombinant and native protein complexes, and more recently, to whole cell lysates or intact unicellular organisms in efforts to identify protein-protein interactions on a global scale. Both traditional non-cleavable and MS-cleavable crosslinkers provide insight into the identification of protein-protein interaction sites, but MS-cleavable crosslinkers are advantageous due to their ability to be cleaved using different types of gas phase fragmentation methods (e.g., CID, HCD, ETD, and EtHCD) and levels of tandem mass spectrometry (e.g., MS2 and MS3) in order to improve identification of cross-linked peptides.

For high resolution analysis of the crosslinked peptides, the recommended LC column for Nanospray Flex source is Acclaim PepMap 100 C18 LC Column (Cat. No. 164940 or 164568). For EASY-Spray source, the recommended LC column is EASY-Spray C18 LC Column (Cat. No. ES800 or ES804).

SureQuant™ AKT Isoform Absolute Quantitation Module (Thermo Scientific™)

The Thermo Scientific SureQuant AKT Isoform Absolute Quantitation Module provides a complete quantitation solution for the analysis of key AKT expression using mass spectrometry (MS).

SureQuant AKT Isoform Absolute Quantitation Module features include:
Complete—contains validated AQUA HeavyPeptide and LightPeptide mixtures, diluent, matrix, system suitability standard including optimized acquisition and analysis methods
Validated—peptides rigorously tested for successful quantitation of each target peptide
Multiplex—able to quantitate AKT1, AKT2, and AKT3 isoforms and phosphorylation of AKT

The AKT signaling pathway serves as a master regulatory pathway for cell growth and proliferation, and AKT plays a vital role in the signaling cascade. Aberrant activation of this pathway promotes survival of tumor cells in many types of cancer. This kit is ideal for monitoring AKT expression levels of the three isoforms and phosphorylation status in a single assay. The ability to simultaneously monitor and quantitate in a single sample is a powerful tool for assessment of normal and abnormal pathway signaling.

The SureQuant AKT Isoform Absolute Quantitation Module provides reagents for absolute quantitation of six unique peptides from AKT using liquid chromatography (LC) and MS. AQUA Ultimate HeavyPeptide mix, AQUA Ultimate LightPeptide mix, and Pierce 6 Protein Digest, equimolar, LC-MS grade reagents are used to generate a standard curve in a matrix, and serial dilutions performed by the user have been optimized to span 2–3 orders of magnitude (200 to 0.03 fmol on column). Standardized methods and data process templates are provided for streamlined data acquisition and analysis.

The Pierce LC-MS/MS System Suitability Standard (7 x 5 Mix) is included in this module to assess dynamic range of LC-MS/MS systems before running the calibration curves and samples.

This kit can be used with SureQuant AKT Isoform IP and MS Sample Preparation Module for enriched multiplex sample preparation. When used together, these modules provide a complete solution for sample preparation and analysis of the target proteins.

Related products:
EASY-Spray LC Column
SureQuant AKT Pathway (Phospho) Multiplex Panel (Absolute Quantitation)
Pierce Trypsin Protease, MS Grade
Pierce LC-MS/MS System Suitability Standard

SureQuant™ TP53 Panel IP and MS Sample Preparation Module (Thermo Scientific™)

The Thermo Scientific SureQuant TP53 Panel IP and MS Sample Preparation Module provides a complete reagent solution for the multiplexed enrichment and sample preparation of TP53 for mass spectrometry (MS).

SureQuant TP53 Isoform IP and MS Sample Preparation Module features include:
Complete—includes all of the reagents necessary for sample lysis, immuno-enrichment, and MS sample preparation
Validated—antibodies validated for specificity and background
Optimized—procedure simplified and condensed to prepare samples after immunoprecipitation in four hours
Multiplex—able to immuno-enrich TP53 and phosphorylated TP53 protein

The tumor suppressor protein TP53 functions as a negative checkpoint regulator for abnormal cell signaling and can induce cell cycle arrest, activate DNA repair proteins, or initiate apoptosis. Mutations in TP53 often result abnormal cell growth and are commonly found in many types of cancer. This module is ideal for multiplexed immuno-enrichment of TP53 protein for use with the SureQuant quantitation modules.

The SureQuant TP53 Panel IP and MS Sample Preparation Module enables highly effective antigen immunoprecipitation (IP) and co-immunoprecipitation (co-IP) for MS analysis. Cells are first lysed and incubated with an IP-MS-verified biotinylated antibody mixture. The antibody-antigen complex is then captured on MS-compatible streptavidin magnetic beads, where non-specific proteins and removed through washing with MS-compatible buffers. The target proteins are eluted using trypsin and are directly compatible with MS sample preparation. The MS sample preparation has been streamlined and can be completed in ~4 hours for same-day MS analysis.

This kit can be used with the either the SureQuant TP53 Panel Absolute Quantitation Module or the SureQuant TP53 Panel Relative Quantitation Module for protein targeted quantitation. When used together, these modules provide a complete solution for sample preparation and analysis of the target proteins.

Related products:
EASY-Spray LC Column
SureQuant AKT Pathway Multiplex Panel (Absolute Quantitation)
Pierce Trypsin Protease, MS Grade
Pierce LC-MS/MS System Suitability Standard

High Select™ HSA/Immunoglobulin Depletion Mini Spin Columns (Thermo Scientific™)

Thermo Scientific High Select HSA/Immunoglobulin Depletion Resin decreases the albumin and antibody components that are abundant in human plasma samples, helping prepare those samples for mass spectrometry or 1D or 2D electrophoresis. The resin is provided here in a convenient single-use mini spin column format for the treatment of 10 µL samples.

Features of HSA/IgG Depletion Resin include:
• Flexible—mini column format is scaled and optimized for treating 10 µL human plasma samples
• Optimized—removes >95% of IgG and >95% of albumin
• Fast—process samples in ~10 minutes
• Economical—cost-effective spin columns are priced for single use
• Reduce variability—one-time use prevents protein carryover and experimental variability

High Select HSA/Immunoglobulin Depletion Resin uses highly specific immobilized anti-HSA and anti-Immuoglobulin antibodies (IgG, IgA, IgM, IgD, and IgE) to remove human serum albumin (HSA) and all major subclasses of immunoglobulins from serum and plasma. The resin is provided in an economical and convenient spin column format specifically designed for one-step processing and single use.

Analysis of human fluids is often complicated by the presence of high concentrations of albumin and IgG that can make up more than 70% of total serum protein. Removal of these proteins is often essential for the study of low-abundant proteins. High Select HSA/Immunoglobulin Depletion Resin can deplete greater than 95% of HSA and 95% of IgG. Samples are loaded in a pre-filled disposable spin column and processed in approximately 10 minutes. The depletion of highly abundant proteins helps enable the detection of low abundant proteins in samples and subsequent identification by mass spectrometry or 1D or 2D gel electrophoresis.

Recommended products
High Select Top14 Abundant Protein Depletion Mini Spin Columns
High Select Top14 Abundant Protein Depletion Resin
TMT10plex Isobaric Label Reagent Set, 3 x 0.8 mg

HCT 116 (+hIGF-1) Control Lysate (1mg/mL) (Thermo Scientific™)

Thermo Scientific HCT 116 (+hIGF-1) Control Lysate is a stimulated cell lysate that has been validated for the expression levels and phosphorylation of key targets in the RAS signaling pathway.

HCT 116 (+hIGF-1) Control Lysate features include:
• Validated for the expression RAS by western blotting and of NRAS, KRAS, and HRAS by mass spectrometry analysis
• Optimized—stimulation and lysis conditions optimized for detection of RAS signaling targets
• Convenient—lysate is ready-to-use for immunoprecipitation, immunoprecipitation to mass spectrometry, and western blotting applications
• Stable for two years at -80°C

The RAS family of GTPase proteins modulates two key signaling pathways involved in cell growth and proliferation: the MAPK and PI3K pathways. Aberrant activation of RAS proteins promotes constitutive activation of cell growth and proliferation that can result in cancer. Mutations in the NRAS, KRAS, and HRAS oncogenes are the most common mutations in human cancer. This lysate has been validated for expression levels of the three isoforms of RAS using mass spectrometry.

The HCT 116 cell line is a human colon cancer epithelial cell line and has been used for proliferation and tumorigenicity studies for colon cancer. The HCT 116 (+hIGF-1) cell lysate is provided as frozen lysate at a standard concentration of 1 mg/mL (500 mg per vial). Lysates are prepared in non-denaturing buffer without loading dyes or dissociating agents (i.e., no 2-mercaptoethanol or dithiothreitol has been added). For western blot analysis, heat lysate to 95°C for 5 minutes. Add reducing agent before heating if desired. The recommended loading volume per lane is 10–20 μL (10–20 µg).

Related products
SureQuant AKT Pathway (Phospho) Multiplex Panel (Absolute Quantitation)
SureQuant RAS Isoform Panel (Absolute Quantitation)
SureQuant TP53 Panel (Absolute Quantitation)

SureQuant™ AKT Pathway Absolute Quantitation Module (Thermo Scientific™)

The Thermo Scientific SureQuant AKT Pathway Absolute Quantitation Module provides a complete quantitation solution for the analysis of key AKT signaling pathway proteins using mass spectrometry (MS).

SureQuant AKT Pathway Absolute Quantitation Module features include:
• Complete—contains validated AQUA HeavyPeptide and LightPeptide mixtures, diluent, matrix, system suitability standard including optimized acquisition and analysis methods
• Validated—peptides rigorously tested for successful quantitation of each target peptide
• Multiplex—able to quantitate AKT, PTEN, IRS1, IGF1-R, GSK3a, GSK3b, PRAS40, mTOR, p70S6K, and TSC2 in a single reaction

The AKT signaling pathway serves as a master regulatory pathway for cell growth and proliferation. Aberrant activation of this pathway promotes survival of tumor cells in many types of cancer. This multiplex kit is ideal for monitoring protein expression of ten key proteins in the AKT signaling pathway (AKT, PTEN, IRS1, IGF1-R, GSK3a, GSK3b, PRAS40, mTOR, p70S6K, and TSC2) in a single assay. The ability to simultaneously monitor and quantitate these targets in a single sample is a powerful tool for assessment of normal and abnormal cell growth.

The SureQuant AKT Pathway Absolute Quantitation Module provides reagents for absolute quantitation of 30 unique peptides from 10 AKT-mTOR signaling pathway target proteins using liquid chromatography (LC) and MS. AQUA Ultimate HeavyPeptide mix, AQUA Ultimate LightPeptide mix, and Pierce 6 Protein Digest, equimolar, LC-MS-grade reagents are used to generate a standard curve in a matrix, and serial dilutions performed by the user have been optimized to span 2–3 orders of magnitude (200 to 0.03 fmol on column). Standardized methods and data process templates are provided for streamlined data acquisition and analysis.

The Pierce LC-MS/MS System Suitability Standard (7 x 5 Mix) is included in this module to assess dynamic range of LC-MS/MS systems before running the calibration curves and samples.

This kit can be used with the SureQuant AKT Pathway IP and MS Sample Preparation Module for enriched multiplex sample preparation. When used together, these modules provide a complete solution for sample preparation and analysis of the target proteins in the pathway.

Related products:
EASY-Spray LC Column
SureQuant AKT Pathway (Phospho) Multiplex Panel (Absolute Quantitation)
Pierce Trypsin Protease, MS Grade
Pierce LC-MS/MS System Suitability Standard

SureQuant™ RAS Isoform Panel (Absolute Quantitation) (Thermo Scientific™)

The Thermo Scientific SureQuant RAS Isoform Panel (Absolute Quantitation) provides a complete reagent and quantitation solution for the enrichment and analysis of NRAS, HRAS, and KRAS isoforms using mass spectrometry (MS).

The SureQuant RAS Isoform Panel (Absolute Quantitation) consists of two modules:
SureQuant RAS Isoform IP and MS Sample Preparation Module includes all of the reagents necessary to immuno-enrich NRAS, KRAS, and HRAS proteins and perform in-solution MS sample preparation from sample lysis through peptide clean-up
SureQuant RAS Isoform Absolute Quantitation Module includes a system suitability standard to monitor MS performance, AQUA Ultimate HeavyPeptide and LightPeptide mixtures for absolute peptide quantitation, and methods for using Thermo Scientific Q Exactive and Fusion Tribrid instrumentation and Nanospray LC instrumentation

SureQuant RAS Isoform Panel features include:
Complete—includes all reagents for successful monitoring of MS system performance, sample preparation, and absolute quantitation of target proteins/peptides
Validated—antibodies, peptides, and control lysate rigorously tested for specificity and successful quantitation of each target peptide
Multiplex—able to quantitate isoforms of RAS
Flexible—modular format allows for immuno-enrichment only, or in combination with relative or absolute quantitation

The RAS family of GTPase proteins modulates two key signaling pathways involved in cell growth and proliferation: the MAPK and PI3K pathways. Aberrant activation of the RAS proteins promotes constitutive activation of cell growth and proliferation that can result in cancer. Mutations in the NRAS, KRAS, and HRAS oncogenes are the most common mutations in human cancer. This kit is ideal for monitoring expression levels of the three isoforms in a single assay. The ability to simultaneously monitor and quantitate in a single sample is a powerful tool for assessment of normal and abnormal pathway signaling.

The SureQuant RAS Isoform IP and MS Sample Preparation Module enables highly effective antigen immunoprecipitation (IP) and co-immunoprecipitation (co-IP) for MS analysis. Cells are first lysed and incubated with an IP-MS-verified biotinylated antibody mixture. The antibody-antigen complex is then captured on MS-compatible streptavidin magnetic beads, where non-specific proteins are removed through washing with MS-compatible buffers. The target proteins are eluted using trypsin and are directly compatible with MS sample preparation. The MS sample preparation has been streamlined and can be completed in ~4 hours for same-day MS analysis.

The SureQuant RAS Isoform Absolute Quantitation Module provides reagents for absolute quantitation of three unique peptides from NRAS, KRAS, and HRAS isoforms using liquid chromatography (LC) and MS. AQUA Ultimate HeavyPeptide mix, AQUA Ultimate LightPeptide mix, and Pierce 6 Protein Digest, equimolar, LC-MS grade reagents are used to generate a standard curve in a matrix, and serial dilutions performed by the user have been optimized to span 2–3 orders of magnitude (200 to 0.03 fmol on column). Standardized methods and data process templates are provided for streamlined data acquisition and analysis.

The Pierce LC-MS/MS System Suitability Standard (7 x 5 Mix) is included in this module to assess dynamic range of LC-MS/MS systems before running the calibration curves and samples.

Together, these modules provide a complete solution for sample preparation and analysis of the target proteins.

Related products:
EASY-Spray LC Column
SureQuant AKT Pathway (Phospho) Multiplex Panel (Absolute Quantitation)
Pierce Trypsin Protease, MS Grade
Pierce LC-MS/MS System Suitability Standard

SureQuant™ TP53 Panel Relative Quantitation Module (Thermo Scientific™)

The Thermo Scientific SureQuant TP53 Panel Relative Quantitation Module provides a complete quantitation solution for the analysis of key TP53 expression and phosphorylation using mass spectrometry (MS).

SureQuant TP53 Panel Relative Quantitation Module features include:
Complete—contains validated AQUA HeavyPeptide mix, diluent, matrix, system suitability standard including optimized acquisition and analysis methods
Validated—peptides rigorously tested for successful quantitation of each target peptide
Multiplex—able to quantitate native and phosphorylated TP53 protein

The tumor suppressor protein TP53 functions as a negative checkpoint regulator for abnormal cell signaling and can induce cell cycle arrest, activate DNA repair proteins, or initiate apoptosis. Mutations in TP53 often result abnormal cell growth and are commonly found in many types of cancer. This kit is ideal for monitoring TP53 expression and phosphorylation status in a single assay. The ability to simultaneously monitor and quantitate in a single sample is a powerful tool for assessment of normal and abnormal pathway signaling.

The SureQuant TP53 Panel Relative Quantitation Module provides reagents for relative quantitation of six unique peptides from AKT using liquid chromatography (LC) and MS. AQUA Ultimate HeavyPeptide mixture is provided for use as a spike-in internal standard for relative quantitation in the multiplexed sample. Standardized methods and data process templates are provided for streamlined data acquisition and analysis.

The Pierce LC-MS/MS System Suitability Standard (7 x 5 Mix) is included in this module to assess dynamic range of LC-MS/MS systems before running the calibration curves and samples.

This kit can be used with the SureQuant TP53 Panel IP and MS Sample Preparation Module for enriched multiplex sample preparation. When used together, these modules provide a complete solution for sample preparation and analysis of the target proteins.

Related products:
EASY-Spray LC Column
SureQuant AKT Pathway (Phospho) Multiplex Panel (Absolute Quantitation)
Pierce Trypsin Protease, MS Grade
Pierce LC-MS/MS System Suitability Standard