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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

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 Pathway (Phospho) Relative Quantitation Module (Thermo Scientific™)

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

SureQuant AKT Pathway (Phospho) 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 phosphorylated 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, and phosphorylation plays a major role in its modulation. 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 phosphorylated 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 pathway signaling.

The SureQuant AKT Pathway (Phospho) Relative Quantitation Module provides reagents for relative quantitation of 30 unique peptides from 10 phosphorylated AKT-mTOR signaling pathway target proteins 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 AKT Pathway (Phospho) 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 phosphorylated 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™ 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

BT-549 Control Lysate (1mg/mL) (Thermo Scientific™)

Thermo Scientific BT-549 Control Lysate is a stimulated cell lysate that has been validated for the expression levels and phosphorylation of key targets in the TP53 signaling pathway.

BT-549 Control Lysate features include:
• Validated for the expression of TP53 and phosphorylated TP53 by western blotting and mass spectrometry analysis
• Optimized—stimulation and lysis conditions optimized for detection of TP53 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 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 lysate has been validated for TP53 expression and phosphorylation.

The BT-549 cell line is a human triple negative epithelial breast cancer cell line derived from mammary gland. The BT-549 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 (Phospho) Absolute Quantitation Module (Thermo Scientific™)

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

SureQuant AKT Pathway (Phospho) 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 phosphorylated 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, and phosphorylation plays a major role in its modulation. 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 phosphorylated 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 pathway signaling.

The SureQuant AKT Pathway (Phospho) Absolute Quantitation Module provides reagents for absolute quantitation of 30 unique peptides from 10 phosphorylated 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 (Phospho) 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 phosphorylated 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™ AKT Pathway (Phospho) Multiplex Panel (Absolute Quantitation) (Thermo Scientific™)

The Thermo Scientific SureQuant AKT Pathway (Phospho) Multiplex Panel (Absolute Quantitation) provides a complete reagent and quantitation solution for the enrichment and analysis of key phosphorylation events of AKT signaling pathway proteins using mass spectrometry (MS).

The SureQuant AKT Pathway (Phospho) Multiplex Panel (Absolute Quantitation) consists of two modules:
SureQuant AKT Pathway (Phospho) IP and MS Sample Preparation Module includes reagents necessary to immuno-enrich phosphorylated AKT-mTOR pathway proteins and perform in-solution MS sample preparation from sample lysis through peptide clean-up
SureQuant AKT Pathway (Phospho) 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 instruments and Nanospray LC instrumentation

SureQuant AKT Pathway (Phospho) Multiplex 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 phosphorylation of ten key proteins in the AKT signaling pathway
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 phosphorylation plays a major role in its modulation. 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 phosphorylated 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 pathway signaling.

The SureQuant AKT Pathway (Phospho) 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, with non-specific proteins 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 Pathway (Phospho) Absolute Quantitation Module provides reagents for absolute quantitation of 30 unique peptides from 10 phosphorylated 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.

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

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

Pierce™ Protein Precipitation Plates (Thermo Scientific™)

Thermo Scientific Pierce Protein Precipitation Plates provide an easy, 96-well format for simple, rapid and automatable precipitation and removal of proteins from plasma, serum and other biological fluid samples.

Features of Protein Precipitation Plates:

• Prepare small molecule samples for HPLC or LC-MS in 10-30 minutes
• 96-well format for automatable protein precipitation and filtration
• Optimized for parallel processing and automation
• Works seamlessly with the Rapid Equilibrium Device (RED) for plasma protein-binding assays
• No acetonitrile leakage for up to four hours
• Leach-free graded hydrophobic frit prevents blockage
• Filtrates are easily collected by vacuum, positive pressure or centrifugation
• Compatible with most organic solvents

These 96-well filtration plates are designed specifically for solvent-based (e.g., acetonitrile) precipitation of proteins and their subsequent removal from fluid samples. The plates are leak-free and retain organic solvent without dripping, even while mixing. Samples are rapidly recovered by applying vacuum or other pressure. The 2 mL well volume allows processing sample volumes of 15 to 400 mL per well, using the recommended precipitation conditions of 3:1 (v/v) solvent-to-serum ratio. The Protein Precipitation Plates are the ideal choice for downstream processing of plasma protein binding assay samples prepared with equilibrium dialysis systems.

The design of Pierce Protein Precipitation Plates provides many advantages over standard protein precipitation methods based solely on either filtration or centrifugation. The plates use a special graded filter system that retains precipitated protein particles without clogging, yet freely allows soluble (nonprecipitated) eluates through the 0.2 µM cut-off membrane. This design maximizes protein removal and ensures the highest possible level of sample recovery.

The study of small molecules and how they behave in complex biofluids (serum, plasma, blood and urine) is an essential part of pharmaceutical, DMPK and metabolomics research. Drugs or other small molecules are routinely monitored for clearance rates, steady-state concentrations, serum-binding equilibrium or in vivo modification. These studies require isolation of target molecules from the protein matrix before analysis. Precipitating proteins with acetonitrile and then removing the precipitate by centrifugation or filtration is the typical isolation process. Although centrifugation to form pellets is inexpensive and reliable, 96-well filtration plates provide an easy format for reproducible, simple, rapid and automatable protein precipitation and sample clarification.

DSBU (Disuccinimidyl Dibutyric Urea) (Thermo Scientific™)

DSBU (disuccinimidyl dibutyric urea), or BuUrBU, is a mass-spectrometry (MS) -cleavable and membrane-permeable cross-linker that contains an amine-reactive N-hydroxysuccinimide (NHS) ester at each end of an 11-atom spacer arm. The cross-linker facilitates analysis of protein structure and complex interactions using mass spectrometry. DSBU has similar reactivity to DSS, but contains a urea 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 DSBU 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 MeroX or XlinkX.

Features of DSBU:
• Reactive group: NHS ester (both ends)
• 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)
• 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 cross-linkers provide insight into the identification of protein-protein interaction sites, but MS-cleavable cross-linkers 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.

References
1. Müller MQ, Dreiocker F, Ihling CH, Schäfer M, Sinz A. Cleavable cross-linker for protein structure analysis: reliable identification of cross-linking products by tandem MS. Anal Chem. 2010 Aug 15;82(16):6958-68.
2. Arlt C, Götze M, Ihling CH, Hage C, Schäfer M, Sinz A. Integrated Workflow for Structural Proteomics Studies Based on Cross-Linking/Mass Spectrometry with an MS/MS Cleavable Cross-Linker. Anal Chem. 2016 Aug 16;88(16):7930-7.

Related products
DSSO (disuccinimidyl sulfoxide), 10 × 1 mg
DSS (disuccinimidyl suberate), 8 × 2 mg
BS3 (bis(sulfosuccinimidyl)suberate), 8 × 2 mg

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).

High Select™ HSA/Immunoglobulin Depletion Resin (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 the samples for mass spectrometry or 1D or 2D electrophoresis.

Features of HSA/Immunoglobulin Depletion Resin include:
• Flexible—provided in bulk format for depletion of user-defined amount of human plasma sample
• Optimized—removes >95% of IgG and >95% of albumin
• Fast—process samples in ~10 minutes
• Economical—cost-effective bulk format

High Select HSA/Immunoglobulin Depletion Resin uses highly specific immobilized anti-HSA and anti-immunoglobulin antibodies (IgG, IgA, IgM, IgD, and IgE) to remove human serum albumin (HSA) and all major subclasses of immunoglobulins from serum or plasma. The resin is provided in an economical bulk format that may be used to fill columns or plates, depending on the sample volumes.

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 resin can deplete greater than 95% of HSA and 95% of IgG. Columns or plates are filled with resin based on the needs of the user, and samples are processed in approximately 10 minutes. The depletion of highly abundant proteins enables 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 Midi Spin Columns
High Select Top14 Abundant Protein Depletion Resin
TMT10plex Isobaric Label Reagent Set, 3 x 0.8 mg

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

The Thermo Scientific SureQuant AKT Isoform Panel (Absolute 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 (Absolute 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 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 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 Absolute Quantitation Module provides reagents for absolute quantitation of six unique peptides from AKT1, AKT2, and AKT3 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 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

High Select™ Top14 Abundant Protein Depletion Resin (Thermo Scientific™)

Thermo Scientific High Select Top14 Abundant Protein Depletion Resin decreases the abundant protein and antibody components of human plasma samples, helping prepare the samples for mass spectrometry or 1D or 2D electrophoresis.

Features of Top14 Abundant Protein Depletion Resin include:
• Flexible—provided in bulk format for depletion of user-defined amount of human plasma sample
• Optimized—remove >95% of IgG and >95% of albumin, plus 12 other abundant proteins (Top 14)
• Fast—process samples in ~10 minutes
• Economical—cost-effective bulk format

High Select Top14 Abundant Protein Depletion Resin uses highly specific immobilized antibodies to remove human serum albumin (HSA), albumin, IgG, IgA, IgM, IgD, IgE, kappa and lambda light chains, alpha-1-acidglycoprotein, alpha-1-antitrypsin, alpha-2-macroglobulin, apolipoprotein A1, fibrinogen, haptoglobin, and transferrin from serum, plasma, or spinal fluids. The resin is provided in an economical bulk format that may be used to fill columns or plates, depending on sample volumes.

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 Top14 Abundant Protein Depletion Resin can deplete greater than 95% of the 14 most abundant proteins in serum. Columns or plates are filled with the resin based on the needs of the user, and samples processed in approximately 10 minutes. The depletion of highly abundant proteins enables the detection of low abundant proteins in samples and subsequent identification by mass spectrometry or 1D or 2D gel electrophoresis.

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 Pathway (Phospho) Multiplex Panel (Relative Quantitation) (Thermo Scientific™)

The Thermo Scientific SureQuant AKT Pathway (Phospho) Multiplex Panel (Relative Quantitation) provides a complete reagent and quantitation solution for the enrichment and analysis of key phosphorylation events of AKT signaling pathway proteins using mass spectrometry (MS).

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

SureQuant AKT Pathway (Phospho) Multiplex 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 phosphorylation of ten key proteins in the AKT signaling pathway
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 phosphorylation plays a major role in its modulation. 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 phosphorylated 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 pathway signaling.

The SureQuant AKT Pathway (Phospho) 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, with non-specific proteins 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 Pathway (Phospho) Relative Quantitation Module provides reagents for relative quantitation of 30 unique peptides from 10 phosphorylated AKT-mTOR signaling pathway target proteins 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 phosphorylated target proteins in the pathway.

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

A549 (+hIGF-1) Control Lysate (1mg/mL) (Thermo Scientific™)

Thermo Scientific A549 (+hIGF-1) Control Lysate is a stimulated cell lysate that has been validated for the expression levels and phosphorylation of key targets in the AKT signaling pathway.

A549 (+hIGF-1) Control Lysate features include:
• Validated for the expression of AKT, PTEN, IRS1, IGF1-R, GSK3a, GSK3b, PRAS40, mTOR, p70S6K, and TSC2 proteins by western blotting and mass spectrometry analysis
• Optimized—stimulation and lysis conditions optimized for detection of AKT 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 AKT signaling pathway serves as a master regulatory pathway for cell growth and proliferation, and phosphorylation plays a major role in its modulation. Aberrant activation of this pathway promotes survival of tumor cells in many types of cancer. This lysate has been validated for expression of ten key proteins in the AKT signaling pathway (AKT, PTEN, IRS1, IGF1-R, GSK3a, GSK3b, PRAS40, mTOR, p70S6K, and TSC2).

The A549 cell line is a human lung carcinoma cell line and is used as a model for non-small lung carcinoma. The A549 (+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).

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