Shop All Mass Spectrometry Sample Preparation Kits

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. Enabled through proprietary MSIA D.A.R.T.'S™ technologies, these workflows have demonstrated pg/mL level detection while providing simultaneous qualitative/quantitative data content from complex biological matrix, such as human plasma. Ideal for biologic and biomarker bio-analysis, MSIA provides innovative enablement in the areas of PKPD, DMPK, ADME, Biotransformation (DAR and payload determination), as well as routine biomarker analysis.

Pierce™ MS-Compatible Magnetic IP Kit, streptavidin Thermo Scientific™

The Thermo Scientific™ Pierce™ MS-Compatible Magnetic IP Kit (Streptavidin) provides mass spectrometry-friendly reagents and an optimized protocol to enable highly effective and efficient immunoprecipitation and co-immunoprecipitation of target antigens upstream of LC-MS analysis.

MS-compatible—reagents directly compatible with in-solution peptide digestion, enriched samples contain minimal detergent residuals detected using LC-MS
Sensitive—procedure successfully enriches low abundance proteins (low ng range)
Low background—binding, wash, and elution buffers optimized to minimize enrichment of background proteins
Reduced antibody contamination—antibody biotinylation reduces contamination in the eluate when compared to other IP methods
Robust—procedure and reagents have been robustly tested with numerous targets to ensure consistent enrichment of low abundant proteins (ng range) with at least 2 peptides identified per protein

The Pierce MS-Compatible Magnetic IP Kit (Streptavidin) uses high-quality Pierce Streptavidin Magnetic Beads and optimized buffers that are compatible with downstream LC-MS sample preparation and analysis. After the immunoprecipitation procedure, the target-enriched elution fraction is directly compatible with in-solution tryptic digestion, eliminating the need for gel purification, detergent removal, or desalting.

This kit has been rigorously validated using numerous target antigens with varying expression levels, including targets previously undetected without enrichment or by western blotting. To support IP and co-IP applications using streptavidin supports, the Pierce Antibody Biotinylation Kit for IP has been specifically designed to enable biotin labeling of antibodies. Additionally, the reagents and procedures have been validated using both manual and automated magnetic separation.

Applications:
• Enrichment of low abundance targets to improve MS sensitivity
• Identification of target proteins that are not detectable by western blotting
• Quantitation of target proteins by LC-MS analysis
• Enrichment of target proteins from plasma samples for MS analysis

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).

Pierce™ MS-Compatible Magnetic IP Kit, protein A/G Thermo Scientific™

The Thermo Scientific™ Pierce™ MS-Compatible Magnetic IP Kit (Protein A/G) provides mass spectrometry-friendly reagents and an optimized protocol to enable highly effective and efficient immunoprecipitation and co-immunoprecipitation of target antigens upstream of LC-MS analysis.

MS-compatible—reagents directly compatible with in-solution peptide digestion, enriched samples contain minimal detergent residuals detected using LC-MS
Sensitive—procedure successfully enriches low abundance proteins (low ng range)
Low background—binding, wash, and elution buffers optimized to minimize enrichment of background proteins
Robust—procedure and reagents have been robustly tested with numerous targets to ensure consistent enrichment of low abundance proteins (ng range) with at least 2 peptides identified per protein

The Pierce MS-Compatible Magnetic IP Kit (Protein A/G) uses high-quality Pierce Protein A/G Magnetic Beads to provide wider flexibility of antibody capture than either Protein A or G alone. The optimized kit components have been formulated to be compatible with downstream LC-MS analysis. After the immunoprecipitation procedure, the target-enriched elution fraction is ready for in-solution tryptic digestion without the need for gel purification, detergent removal, or desalting.

This kit has been rigorously validated using numerous target antigens with varying expression levels, including targets previously undetected without enrichment or by western blotting. Additionally, the reagents and procedures have been validated using both manual and automated magnetic separation.

Applications:
• Enrichment of low abundance targets to improve MS sensitivity
• Identification of target proteins that are not detectable by western blotting
• Quantitation of target proteins by LC-MS analysis
• Screening of antibodies for IP-MS
• Enrichment of target proteins upstream of targeted analysis (SRM)

High Select™ Top14 Abundant Protein Depletion Mini Spin Columns 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. The resin is provided here in a convenient single-use mini spin column format for the treatment of 10 µL samples.

Features of Top14 Abundant Protein Depletion Resin include:
• Flexible—mini spin column format is scaled and optimized for treating 10 µL human plasma samples
• Optimized—removes >95% of IgG and >95% of albumin, plus 12 other abundant proteins (Top 14)
• 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 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 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 Top14 Abundant Protein Depletion Resin can deplete greater than 95% of the 14 most abundant proteins in serum. Samples are loaded in a pre-filled disposable spin column and 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™ C18 Spin Tips Thermo Scientific™

The Thermo Scientific Pierce C18 Spin Tips enable fast and efficient capture, concentration, desalting, and elution of up to 10 µg peptides per 20 µL sample for mass spectrometry (MS) analysis.

Pierce C18 Spin Tips are 20 µL-capacity pipette tips with accompanying adaptors for microcentrifuge sample processing. The tips contain a C18 reversed-phase sorbent that minimizes flow resistance and provides excellent binding and recovery characteristics at a wide range of peptide concentrations upstream of matrix-assisted laser desorption ionization (MALDI) or nanoelectrospray ionization techniques. Sample clean-up with C18 resin significantly improves protein analysis results by removing urea, salts, and other contaminants before mass spectrometry (MS). Each Pierce C18 Spin Tip has a 20 µL volume capacity with a 10 µg peptide-binding capacity.

Features of C18 Spin Tips:

Rapid—C18 fast-flow tips have low resistance and improved flow characteristics compared to other commercially available tips
High capacity—up to 10 µg peptide per 20 µL solution
Convenient—spin tips come with tip adaptors for easy centrifugation
Cleaner sample—device design filters out particulates that can cause autosampler and column clogging

Applications:
• Clean up of peptides after protein digestion
• Peptide sample concentration and desalting
• Sample preparation for MALDI peptide analysis
• Sample processing in proteomics workflows

Pierce C18 Spin Tips offer excellent flow properties with a high-efficiency C18 sorbent for fast wetting, loading, washing and eluting. Sample is simply loaded in prepared tip and washed and eluted using multiple centrifugation steps. The procedure is simple and requires less than 5 minutes to process protein digests, strong cation exchange fractions and other protein and peptide samples for mass spectrometric (MS) analyses. The unique tip design also removes any particulates in the sample, eliminating clogging of the auto sampler or column upstream of the mass spectrometer.

High Select™ Top14 Abundant Protein Depletion Midi Spin Columns 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. The resin is provided here in a convenient single-use midi spin column format for the treatment of 100 µL samples.

Features of Top14 Abundant Protein Depletion Resin include:
• Flexible—midi spin column format is scaled and optimized for treating 100 µL human plasma samples
• Optimized—removes >95% of IgG and >95% of albumin, plus 12 other abundant proteins (Top 14)
• 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 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 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 Top14 Abundant Protein Depletion Resin can deplete greater than 95% of the 14 most abundant proteins in serum. Samples are loaded in a pre-filled disposable spin column and 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™ Lys-C Protease, MS Grade Thermo Scientific™

Thermo Scientific™ Pierce™ Lys-C Protease, MS Grade is a highly purified native endoproteinase validated for maximum activity and stability in proteomic applications.

Features of Lys-C Protease, MS Grade include:
Enhanced digestion—use in tandem with trypsin to decrease tryptic missed cleavages
Increased sequence coverage—better protein characterization results from overlapping peptides with complementary chromatographic, ionization, and fragmentation properties
Versatility—effective enzyme activity under highly denaturing conditions (e.g., 8 M urea)
C-terminal lysine cleavage specificity—at least 90% for a complex protein sample
Stability—provided in a lyophilized format

Product details
This is a mass spectrometry (MS)-grade serine protease isolated from Lysobacter enzymogenes. Lys-C protease has high activity and specificity for lysine residues, resulting in larger peptides and less sample complexity than trypsin (i.e., fewer peptides). Unlike trypsin, Lys-C protease can cleave lysines followed by prolines, making it ideal for sequential protein digestion followed by trypsin to decrease missed cleavages. These unique Lys-C protease properties ensure high digestion efficiency when used alone or followed by tryptic digestion. Additionally, Lys-C prototypic peptides typically have higher charge states, making it an enzyme of choice for use with ETD fragmentation.

Lys-C protease is commonly used in phosphopeptide enrichment workflows because it generates peptides with primary amines at both the N- and C-terminus, allowing the fragments to be double-labeled with amine-reactive isobaric tags. This results in enhanced peptide ionization and improved limits of quantitation since more fragment ions can be re-isolated during MS3 acquisition. This enzyme can be used for in-solution or in-gel digestion workflows to produce peptides for LC-MS/MS protein identification. This Lys-C enzyme is packaged lyophilized (20 µg or 100 µg quantities).

The endoproteinase Lys-C specifically hydrolyzes proteins at the carboxyl side of lysine. Efficient protein digestion can be completed in 2 hours at 37°C. Lys-C protease remains active in highly denaturing conditions such as 8 M urea, 2 M guanidine-HCl, 1% SDS, 2% CHAPS, and 40% acetonitrile and functions well within pH 7–9 (maximal activity at pH 8). This lyophilized enzyme has a mass of 30 kDa and is stable for 1 year when stored at –20°C.

Applications
• Improved sequence coverage of protein digests
• De novo sequencing
• Epigenetic studies
• In-gel and in-solution digestion of proteins

Pierce™ High pH Reversed-Phase Peptide Fractionation Kit Thermo Scientific™

Thermo Scientific™ Pierce™ High pH Reversed-Phase Peptide Fractionation Kit increases protein identification from LC/MS analysis through orthogonal peptide fractionation of complex peptide samples.

Easy-to-use—resin provided in single-use spin column format
Improved protein identifications—protein identifications increased by ≥50% when compared to unfractionated samples
Reproducible—elution profiles and fractional resolution vary by less than 20%
Optimized—robust procedure for maximal protein identification and peptide recovery while minimizing fractional overlap
Compatible—reagents validated with a variety of complex samples, including TMT™-labeled peptides

In order to enable deep proteome sequencing, it is often necessary to reduce the sample complexity by fractionation in an orthogonal dimension prior to LC/MS analysis. The Pierce High pH Reversed-Phase Peptide Fractionation Kit utilizes high pH reversed-phase chromatography to separate peptides by hydrophobicity and provides excellent orthogonality to low pH reversed-phase LC-MS gradients.

The kit has been designed to improve protein identification through the use of a proprietary reversed-phase resin in an easy-to-use spin column format with a highpH fractionation protocol. In contrast to strong cation exchange (SCX) fractionation, the high-pH reversed-phase fractions do not require an additional desalting step before LC/MS analysis.

The High pH Reversed-phase Peptide Fractionation Kit includes a high pH buffer (0.1% triethylamine) and twelve spin columns containing pH-resistant reversed-phase resin. Each reversed-phase fractionation spin column enables fractionation of 10–100 µg of peptide sample using a microcentrifuge.

Native, phosphorylated, Tandem Mass TagTM (TMTTM)-labeled, and other complex peptide mixture samples can be fractionated using this kit. Combining the search results generated by the individual fractions helps improve protein sequence coverage and increases the number of identified proteins relative to unfractionated samples.

Applications:
• Reducing sample complexity to identify targets of interest
• Conducting systems biology studies
• Reducing sample complexity to improve quantitation studies

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)

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™ Graphite Spin Columns Thermo Scientific™

Thermo Scientific Pierce Graphite Spin Columns improve phosphopeptide analysis by efficiently binding hydrophilic peptides and efficiently removing urea, salts and other contaminants before mass spectrometry analysis.

Features of Graphite Spin Columns:

• Convenient spin format for parallel processing of multiple samples
• High-binding capacity with excellent recovery of up to 100 µg of hydrophilic peptides per column
• Porous graphite resin for cleaning up phosphopeptide samples before MS analysis

Pierce Graphite Spin Columns enable fast and efficient capture, concentration, desalting and elution of hydrophilic peptides in less than 10 minutes. These columns are ideal for improving mass spectrometric (MS) analyses of samples from protein digests), strong-cation exchange fractions, and enriched phosphopeptides eluted from titanium dioxide and immobilized metal affinity chromatography (IMAC) columns and tips.

The Pierce Graphite Spin Columns improve phosphopeptide analysis by efficiently binding hydrophilic peptides and efficiently removing urea, salts and other contaminants before MS analysis. The C18 resins and C18 tips that are commonly used to desalt peptides are excellent for use with hydrophobic peptides but do not efficiently capture hydrophilic peptides, like phosphopeptides, resulting in enrichment of only hydrophobic fragments. The Pierce Graphite Spin Columns address this issue and are ideal for matrix-assisted laser desorption ionization (MALDI) or nanoelectrospray ionization techniques.

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

The Thermo Scientific SureQuant RAS Isoform Panel (Relative 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 (Relative 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 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 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 Relative Quantitation Module provides reagents for relative quantitation of three unique peptides from NRAS, KRAS, and HRAS 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™ Trypsin/Lys-C Protease Mix, MS-Grade Thermo Scientific™

Thermo Scientific Pierce Trypsin/Lys-C Protease Mix, MS-Grade, is a mass spectrometry (MS)-grade serine endoproteinase mixture of trypsin and LysC that can be used for concurrent digestion of proteins for more efficient digestion than with trypsin alone.

Features of Pierce Trypsin/Lys-C Protease Mix, MS-Grade, include:
Enhanced digestion—enzyme combination reduces tryptic missed cleavages
Convenient—trypsin and LysC proteases provided at an optimized ratio for digestion in a combined-use format
Exceptional selectivity—trypsin has >95% C-terminal lysine and arginine specificity; LysC has >90% C-terminal lysine cleavage specificity
Stable—enzyme mixture provided in a lyophilized format

Protein characterization, identification, and quantification by MS begins with efficient, reproducible protein digestion. Although trypsin is routinely used for protein digestion, this protease alone is not sufficient to fully digest proteins at the carboxyl-end of lysine and arginine residues. Therefore, Lys-C protease is commonly combined with trypsin to sequentially digest proteins with fewer missed cleavages. Pierce Trypsin/Lys-C Protease Mix is a lyophilized mixture of trypsin and LysC proteases that has been optimized to improve digestion efficiency of proteins. It is provided in flexible formats of 20 µg, 5 x 20 µg, or 100 µg, and is also included in the EasyPep Mini MS Sample Prep Kit.

The MS-grade trypsin protease in this mix is derived from porcine pancreatic extracts and has been TPCK-treated to eliminate chymotryptic activity and methylated to improve stability during digestion. The MS-grade Lys-C protease is a highly purified native enzyme from <Lysobacter enzymogenes. Unlike trypsin, Lys-C can cleave lysines followed by prolines, making it ideal for use in combination with other proteases for optimal protein digestion. When used as a mixture, digestion can be completed in as little as 1.5–3 hours or up to overnight, depending on enzyme to protein ratio. This lyophilized enzyme mixture is stable for one year when stored at -20°C.

Related products
EasyPep Mini MS Sample Prep Kit
Pierce Colorimetric Peptide Quantitation Assay
Chloroacetamide, No-Weigh Format

Pierce™ Mass Spec Sample Prep Kit for Cultured Cells Thermo Scientific™

The Thermo Scientific Pierce Mass Spec Sample Prep Kit for Cultured Cells is an easy-to-use, comprehensive kit for preparation of clean peptide mixtures from cultured cells for mass spectrometry (MS) analysis.

Features of the Mass Spec Sample Prep Kit:

Complete—includes all reagents, a digestion indicator control, proteases and an optimized protocol needed to process up to 20 samples
Simple—user friendly kit can provide reproducible results even for non-expert MS analysts
Flexible—can be adapted to handle sample sizes between 10 to 200 µg
High yield—total protein yield from 1 million cells is greater than 100 µg
Optimized—cysteine reduction and alkylation are 100% with less than 1% over alkylation of non-cysteine residues
Efficient - percentage of missed cleavages is less than 10%
Compatible—final preparation is ready for direct MS analysis and other downstream applications, including mass-tag labeling

The Pierce Mass Spec Sample Prep Kit for Cultured Cells contains all the necessary reagents and enzymes to prepare up to 20 samples (1 million cells each) for MS analysis. The simple and robust workflow uses a lysis protocol that generates approximately 100 µg of protein per sample and includes a control digestion indicator to monitor the efficiency of the two-step enzymatic digestion protocol. The kit procedure, which includes a Digestion Indicator as an internal protein/peptide control, produces consistently and reproducibly clean peptide mixtures for protein identification.

Includes:
Kit contains proprietary lysis and digestion buffers, digestion indicator, DTT, iodoacetamide, Lys-C, trypsin

Sample prep for mass spectrometry (MS, mass spec) remains one of the largest bottlenecks associated with MS analysis, and consistent and reproducible sample preparation can make the difference between a successful analysis and a failed outcome. Current sample prep protocols are primarily homebrew and can be highly variable, making data analysis and interpretation difficult. The Mass Spec Sample Prep Kit for Cultured Cells provides researchers with all the necessary tools to generate consistent and reproducible protein digests that are directly compatible with LC-MS workflows. The digests do not require further processing, such as C18 clean up or detergent removal.

The included Pierce Digestion Indicator, which can also be purchased separately, is a non-mammalian recombinant protein (26kDa) with five signature peptides for use in determining the digestion efficiency and reproducibility across multiple samples. The protein sequence and recommended peptides to monitor across samples are supplied with the product instructions.

 

 

 

More Product Data
A versatile mass spectrometry sample preparation procedure for complex protein samples

Related Products
Pierce™ Digestion Indicator for Mass Spectrometry
Results per page
    spinner