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Pierce™ Asp-N Protease, MS Grade (Thermo Scientific™)

Thermo Scientific Asp-N Protease, MS Grade, is a highly specific endoproteinase used to improve sequence coverage in mass spectrometry protein identification applications.

Features of MS Grade Asp-N Protease:

• Complementary to tryptic digests—hydrolyzes proteins specifically at the amino side of aspartate and cysteic acid residues
• Increased sequence coverage—better protein characterization results from overlapping peptides with complementary chromatographic, ionization and fragmentation properties
• High specific activity—greater than 20,000 units/mg protein
• N-terminal arginine cleavage specificity—at least 90% for a complex protein sample
• Stable—provided in a lyophilized format

Product Details
This Asp-N is a mass spectrometry (MS)-grade zinc metalloproteinase derived from a mutant strain of Pseudomonas fragi and requires a trace amount of zinc for activity. Asp-N can be used alone or in parallel with trypsin or other proteases to produce protein digests for peptide mapping and protein sequencing. Asp-N protease is suitable for either in-solution or in-gel digestion workflows. This Asp-N enzyme is packaged lyophilized (2 µg).

The endoproteinase AspN cleaves primarily at amino side of aspartate residues and cysteic acid residues that result from the oxidization of cysteine residues, generating a limited number of peptide fragments. Cleavage can also occur at glutamic residues; however, the rate of cleavage at the glutamyl residues is significantly lower than the rate of cleavage at the aspartic acid residues. AspN can efficiently digest protein in 2-20 hours at 37°C. AspN remains active under denaturing conditions such as 1M urea, 2M guanidine·HCl, 0.1% SDS, 2% CHAPS and 10% acetonitrile with optimal activity in the pH range of 6-8. This lyophilized enzyme has a mass of 27 kDa and is stable for 1 year when stored at -20°C.

Applications
• Improved sequence coverage of protein digests
• In-solution digestion of proteins
• In-gel digestion of proteins

Pierce™ Trypsin Protease, MS Grade (Thermo Scientific™)

Thermo Scientific Pierce Trypsin Protease, MS Grade, is highly purified porcine trypsin that has been chemically modified for maximum activity and stability in proteomic applications.

Pierce Trypsin Protease, MS Grade, is a serine protease derived from porcine pancreatic extracts. The enzyme is TPCK-treated to eliminate chymotryptic activity and methylated to improve stability during protein digestion.

Features of Trypsin Protease, MS Grade:

Exceptional selectivity—cleaves at the carboxyl side of lysine and arginine residues with greater than 95% specificity
High purity—no detectable chymotrypsin activity
Enhanced stability—chemically modified for reduced autolytic activity

Applications:
• In-gel digestion of proteins from 1-D or 2-D gels
• In-solution tryptic digestion of proteins

Trypsin is a serine protease that specifically cleaves at the carboxyl side of lysine and arginine residues. The selectivity of this enzyme is critical for reproducible protein digestion and mass spectrometry-based protein identification. Since chymotrypsin co-purifies with trypsin derived from natural sources, Pierce Trypsin Protease has been treated with TPCK to eliminate chymotryptic activity, improving digestion specificity. Native trypsin is also subject to autolysis which can reduce enzyme stability and efficiency. To reduce autolytic degradation, Pierce Trypsin Protease is chemically modified by methylation, yielding a highly active and more stable form of the enzyme.

In addition to possessing high specific activity and being resistant to autolytic digestion, Pierce Trypsin Protease can tolerate commonly used partially denaturing conditions, such as 0.1% SDS, 1M urea and 10% acetonitrile. Pierce Trypsin Protease is most active in pH ranges pH 7 to 9 and can be reversibly inactivated at pH < 4. Both the liquid and lyophilized forms of the enzyme are stable for > 1 yearr when stored at -20°C.

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Pierce™ Trypsin Protease, MS Grade (Thermo Scientific™)

Thermo Scientific Pierce Trypsin Protease, MS Grade, is highly purified porcine trypsin that has been chemically modified for maximum activity and stability in proteomic applications.

Pierce Trypsin Protease, MS Grade, is a serine protease derived from porcine pancreatic extracts. The enzyme is TPCK-treated to eliminate chymotryptic activity and methylated to improve stability during protein digestion.

Features of Trypsin Protease, MS Grade:

Exceptional selectivity—cleaves at the carboxyl side of lysine and arginine residues with greater than 95% specificity
High purity—no detectable chymotrypsin activity
Enhanced stability—chemically modified for reduced autolytic activity

Applications:
• In-gel digestion of proteins from 1-D or 2-D gels
• In-solution tryptic digestion of proteins

Trypsin is a serine protease that specifically cleaves at the carboxyl side of lysine and arginine residues. The selectivity of this enzyme is critical for reproducible protein digestion and mass spectrometry-based protein identification. Since chymotrypsin co-purifies with trypsin derived from natural sources, Pierce Trypsin Protease has been treated with TPCK to eliminate chymotryptic activity, improving digestion specificity. Native trypsin is also subject to autolysis which can reduce enzyme stability and efficiency. To reduce autolytic degradation, Pierce Trypsin Protease is chemically modified by methylation, yielding a highly active and more stable form of the enzyme.

In addition to possessing high specific activity and being resistant to autolytic digestion, Pierce Trypsin Protease can tolerate commonly used partially denaturing conditions, such as 0.1% SDS, 1M urea and 10% acetonitrile. Pierce Trypsin Protease is most active in pH ranges pH 7 to 9 and can be reversibly inactivated at pH < 4. Both the liquid and lyophilized forms of the enzyme are stable for > 1 yearr when stored at -20°C.

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Pierce™ Glu-C Protease, MS Grade (Thermo Scientific™)

Thermo Scientific Glu-C Protease, MS Grade, is a highly specific endoproteinase use to improve sequence coverage in mass spectrometry protein identification applications.

Features of Glu-C Protease, MS Grade:

• Complementary to tryptic digests—hydrolyzes proteins specifically at the carboxyl side of glutamic acids
• Increased sequence coverage—better protein characterization results from overlapping peptides with complementary sequence coverage and chromatographic, ionization and fragmentation properties
• High specific activity—greater than 500 units/mg protein
• N-terminal arginine cleavage specificity—at least 90% for a complex protein sample
• Stable—provided in a lyophilized format

Product Details
This Glu-C is a mass spectrometry (MS)-grade serine protease isolated from Staphylococcus aureus. Glu-C can be used alone or alongside trypsin or other proteases to produce complementary protein digests for peptide mapping and protein sequencing. Glu-C protease is suitable for either in-solution or in-gel digestion workflows. This Glu-C enzyme is packaged lyophilized (5 x 10 µg).

The endoproteinase Glu-C, also referred to as V-8 Protease, specifically cleaves the carboxyl side of glutamic residues when reactions are carried out in ammonium bicarbonate and ammonium acetate buffers, generating a limited number of peptide fragments. Cleavage can also occur at both glutamic and aspartic residues in phosphate buffers. Glu-C can efficiently digest protein in 5-18 hours at 37°C. Glu-C Protease remains active under denaturing conditions such as 2M urea, 1M guanidine·HCl, 0.1% SDS, 2% CHAPS and 20% acetonitrile. Glu-C activity is optimal at pH 8. This lyophilized enzyme has a mass of 27 kDa and is stable for 1 year when stored at -20°C.

Applications
• Improved sequence coverage of protein digests
• In-solution digestion of proteins
• In-gel digestion of proteins

In-Gel Tryptic Digestion Kit (Thermo Scientific™)

Thermo Scientific Pierce In-Gel Tryptic Digestion Kit provides a complete set of reagents for digesting and recovering peptide fragments from approximately 150 stained protein bands excised from SDS-PAGE (polyacrylamide gel plugs).

Features of the kit:

Convenient—includes all necessary reagents for destaining coomassie- or fluorescent dye-stained proteins, reduction and alkylation of cystines, and tryptic digestion
Robust—the procedure and reagents produce reliable digestions and data generation using a wide range of conditions without optimization
Accurate—contains highly purified and modified MS-grade trypsin that shows no chymotryptic activity and minimal autolytic activity

The In-Gel Tryptic Digestion Kit provides a complete set of reagents for performing approximately 150 digestions on colloidal coomassie- or fluorescent dye-stained protein bands. The kit includes modified porcine trypsin, destaining buffers, reduction reagents, alkylation reagents and digestion buffers along with detailed and simple instructions. Each component and step has been optimized and balanced to produce complete, accurate and clean digests using a variety of conditions for dependable proteomics analysis of peptide profiles by mass spectrometry (MS).

Accurate identification and analysis of proteins is a central component of proteomic strategies for studying cellular functions and processes. Mass spectrometers are essential tools for these studies as they provide a high level of sensitivity and mass accuracy. However, this sensitivity and accuracy can be obtained only if proper sample preparation is performed. Two key sample preparation steps are the fragmentation of proteins into peptides and subsequent concentration and clean up of peptides. The In-Gel Tryptic Digestion Kit accomplishes the first of these steps. Thermo Scientific Pierce C-18 Columns accomplish the second step.

Pierce™ Chymotrypsin Protease (TLCK treated), MS Grade (Thermo Scientific™)

Thermo Scientific Chymotrypsin Protease, MS Grade, is a purified native endoproteinase validated for maximum activity and selectivity in proteomic applications.

Features of Chymotrypsin Protease, MS Grade:

• Increased sequence coverage—cleaves at the carboxyl side of tyrosine, phenylalanine, tryptophan and leucine
• Better specificity—treated with TLCK to eliminate trypsin activity
• High activity—greater than 45 units/mg protein

This Chymotrypsin is a mass spectrometry (MS)-grade serine protease isolated from bovine pancreatic extracts. This chymotrypsin is TLCK-treated to eliminate tryptic activity. The selectivity of this enzyme is important for reproducible protein digestion and mass spectrometry-based protein identification. Chymotrypsin-digestion typically generates a larger number of shorter peptides compared to trypsin. This mass spectrometry-grade Chymotrypsin is packaged lyophilized (convenient 4 x 25 µg).

Applications
• In-solution digestions
• In-gel digestion of proteins from 1-D or 2-D gels

Product Details
The endoproteinase chymotrypsin specifically cleaves at the carboxyl side of tyrosine, phenylalanine, tryptophan and leucine. Two predominant forms of chymotrypsin, A and B, are found in equal amounts in bovine pancreas. They are similar proteins (80% homology), but have different proteolytic characteristics. Both forms of chymotrypsin are present in Thermo Scientific Chymotrypsin. Since trypsin may co-purify with Chymotrypsin derived from natural sources, Thermo Scientific Chymotrypsin has been treated with TLCK to eliminate potential tryptic activity, improving digestion specificity. Chymotrypsin can tolerate mild denaturing conditions, such as 0.1% SDS, 2M urea, 2M guanidine·HCl, 1% CHAPS, and 30% acetonitrile with optimal activity in the pH range 7.5 to 8.5. This lyophilized enzyme has a mass of 25 kDa and is stable for 1 year when stored at -20°C.

Pierce™ Lys-C Protease, MS Grade (Thermo Scientific™)

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

Features of MS Grade Lys-C Protease:

• 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
• Versatile—effective enzyme activity under highly denaturing conditions (e.g., 8M urea)
• C-terminal lysine cleavage specificity—at least 90% for a complex protein sample
• Stable—provided in a lyophilized format

Product Details
This Lys-C is a mass spectrometry (MS)-grade serine protease isolated from Lysobacter enzymogenes. Lys-C 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 can cleave lysines followed by prolines, making it ideal for sequential protein digestion followed by trypsin to decrease missed cleavages. These unique Lys-C 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 is commonly 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 LysC specifically hydrolyzes proteins at the carboxyl side of lysine. Efficient protein digestion can be completed in 2 hours at 37°C. LysC remains active in highly denaturing conditions such as 8M urea, 2M 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

In-Solution Tryptic Digestion and Guanidination Kit (Thermo Scientific™)

The Thermo Scientific Pierce In-Solution Tryptic Digestion and Guanidination Kit contains proteomics-grade trypsin, buffers and reagents to alkylate and digest proteins and then guanidinate the peptide fragments for mass spectrometry.

Features of the kit:

Optimized trypsin digestion—produce complete digests of protein ranging from 0.025 to 10 µg
Optimized guanidination—reaction produces little to no N-terminal modification
Convenient—kit includes reagents for digestion, reduction, alkylation and guanidination.
Quick—protein can be digested and guanidinated all in one day

Accurate identification of proteins and analysis of post-translational modifications by mass spectrometry require accurate and complete protein digestion and peptide modification. The In-Solution Tryptic Digestion and Guanidination kit provides an optimized procedure and reagents for approximately 90 digests, each containing 0.025 to 10 µg of protein.

Trypsin specifically cleaves peptide bonds at the carboxyl side of arginine and lysine residues. The signals from the arginine-containing peptides are generally stronger, due to the more basic side chain. To enhance overall ionization, guanidination is necessary to convert lysines to homoarginines. The guanidination reaction is specific for the amine of lysine but may occur minimally at the amine of the peptide's N-terminus. This derivatization leads to an increase in the intensity of the lysine containing peptides and improved sequence coverage overall.

Pierce™ LysN Protease, MS Grade (Thermo Scientific™)

Thermo Scientific Pierce LysN Protease, MS Grade, is highly purified native Lys-N protease that has been validated for maximum activity and stability in proteomic applications.

Features of LysN Protease, MS Grade:

Thermostable – provides better digestion at higher temperatures (e.g., 50°C) in less time
Complementary to tryptic digests cleaves the opposite side of lysine from trypsin
High purity – no additional (off-target) protease activity detected
N-terminal lysine cleavage specificity – at least 90% for a complex protein sample
Versatile – enzyme is effective at many temperatures and in denaturing conditions
Stable – provided in a lyophilized format

Thermo Scientific Pierce LysN is a mass spectrometry (MS)-grade zinc metalloprotease derived from Grifola frondosa. The native Pierce LysN Protease has been highly purified to improve stability, specific activity, and cleavage selectivity. Unlike trypsin, LysN protease cleaves at the amino-terminus of lysine residues. As a result, the peptides generated by LysN are longer than those generated by trypsin and have more prevalent charged amino terminal peptide fragments. Additionally, LysN protease is more promising for epigenetic MS applications than LysC or trypsin because it is capable of cleaving methylated lysines [1]. The thermostability and chemical compatibility with denaturants make LysN protease ideal for digestion of complex protein samples for “shotgun” proteomics.

Applications:
• Improved sequence coverage of protein digests
• De novo sequencing with collision induced dissociation (CID)
• Epigenetic studies
• In-solution digestion of proteins

LysN protease is active over a wide range of temperatures and denaturing conditions. Efficient protein digestion can be completed in 2 hours at 50°C or 4 hours at 37°C, however, digestion is possible at both lower and higher temperatures. LysN also remains active under moderate denaturing conditions including 0.1% SDS, 6M urea or heating to 70°C. Maximal LysN activity occurs at pH 7 to 9. The lyophilized enzyme is stable for 2 years when stored at -20°C, and reconstituted stock solutions of LysN are stable at -80°C for 2 years or -20°C for one year without significant loss in activity.

Pierce™ Digestion Indicator for Mass Spectrometry (Thermo Scientific™)

The Thermo Scientific Pierce Digestion Indicator for Mass Spectrometry is a non-mammalian recombinant protein (26kDa) with five signature peptides for use in determining the digestion efficiency and reproducibility across multiple samples. This product is sufficient for production of five signature peptides upon digestion for mass spectrometry.

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

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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.

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Pierce™ Trypsin Protease, MS Grade, Frozen (Thermo Scientific™)

Thermo Scientific Pierce Trypsin Protease, MS Grade, is highly purified porcine trypsin that has been chemically modified for maximum activity and stability in proteomic applications.

Pierce Trypsin Protease, MS Grade, is a serine protease derived from porcine pancreatic extracts. The enzyme is TPCK-treated to eliminate chymotryptic activity and methylated to improve stability during protein digestion.

Features of Trypsin Protease, MS Grade:

Exceptional selectivity—cleaves at the carboxyl side of lysine and arginine residues with greater than 95% specificity
High purity—no detectable chymotrypsin activity
Enhanced stability—chemically modified for reduced autolytic activity

Applications:
• In-gel digestion of proteins from 1-D or 2-D gels
• In-solution tryptic digestion of proteins

Trypsin is a serine protease that specifically cleaves at the carboxyl side of lysine and arginine residues. The selectivity of this enzyme is critical for reproducible protein digestion and mass spectrometry-based protein identification. Since chymotrypsin co-purifies with trypsin derived from natural sources, Pierce Trypsin Protease has been treated with TPCK to eliminate chymotryptic activity, improving digestion specificity. Native trypsin is also subject to autolysis which can reduce enzyme stability and efficiency. To reduce autolytic degradation, Pierce Trypsin Protease is chemically modified by methylation, yielding a highly active and more stable form of the enzyme.

In addition to possessing high specific activity and being resistant to autolytic digestion, Pierce Trypsin Protease can tolerate commonly used partially denaturing conditions, such as 0.1% SDS, 1M urea and 10% acetonitrile. Pierce Trypsin Protease is most active in pH ranges pH 7 to 9 and can be reversibly inactivated at pH < 4. Both the liquid and lyophilized forms of the enzyme are stable for > 1 yearr when stored at -20°C.

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