Product Class: Other

α-Lytic Protease
cloned at NEB 37 95 Heat

Product Introduction

P8113 Cutsite

α-Lytic Protease (aLP) cleaves after Threonine (T), Alanine (A), Serine (S) and Valine (V) residues. Its specificity makes it an orthogonal and alternative protease to others commonly used in proteomics applications, including trypsin and chymotrypsin.

  • Analyze complex proteomes
  • Suitable for both in-gel and solution digests
  • Optimal activity and stability for up to 24 months
  • Ideal for digestion of proteins for proteomic analysis by mass spectrometry
Catalog # Size Concentration
P8113S 20.0 µg 0.4 mg/ml
P8113L 100.0 µg 0.4 mg/ml

Product Information

Description

α-Lytic Protease (aLP) cleaves after Threonine (T), Alanine (A), Serine (S) and Valine (V) residues. Its specificity makes it an orthogonal and alternative protease to others commonly used in proteomics applications, including trypsin and chymotrypsin. Peptides generated by aLP are of similar average length to those of Trypsin.

Figure 1.  α-Lytic Protease can be used alone or in combination with other proteases to yield increased sequence coverage.



Comparison of sequence coverage for three protein standards after parallel digestion using Trypsin (blue) and a-Lytic Protease (gold). The combined data set (grey) results in overlapping peptides and increased sequence coverage.

Product Source

Purified from Lysobacter enzymogenes
This product is related to the following categories:
Proteases Products,
Proteome Analysis Products,
This product can be used in the following applications:
Protein Analysis Tools,
Protein Digestion,
Proteomics

Reagents Supplied

Reagents Supplied

The following reagents are supplied with this product:

NEB # Component Name Component # Stored at (°C) Amount Concentration

Properties & Usage

Storage Buffer

10 mM sodium acetate
pH 5 @ 25°C

Heat Inactivation

95°C

Molecular Weight

Apparent: 19.8 kDa

Features

  • Digestion of proteins for proteomic analysis by mass spectrometry
  • Protein and peptide identification
  • Site-specific identification and quantitation of endogenous SUMO modifications under native conditions(1). 
  • Cleavage after aliphatic amino acids resulting in increased sequence coverage of membrane proteins 

Product Notes

  1. α-Lytic Protease is stable for at least 2 years at –20°C. No loss of activity is observed after 10 freeze-thaw cycles.
  2. To avoid autolysis, α-Lytic Protease should be stored long term in 10 mM Sodium Acetate pH 5.0. 
  3. α-Lytic Protease is active in a variety of mass spectrometry compatible buffers including ammonium bicarbonate, Tris-HCl, and HEPES. The optimal pH range for α-Lytic Protease digestion is pH 7.5 - 8.5.
  4. Enzyme activity is stimulated in up to 0.1% sodium deoxycholate(2). 
  5. Enzyme activity is inhibited by 1.0% sodium deoxycholate (~60% active); 0.1% SDS (~50 active); 1% SDS (~40% active); 1 M guanidine hydrochloride (~20% active); 4 M guanidine hydrochloride (no activity); Serine protease inhibitors, such as PMSF (no activity)(2).
  6. α-Lytic Protease is compatible with both in-solution and in-gel digestion methods. Digestion with α-Lytic Protease yields increased peptide recovery for standard and crushed gel conditions of large molecular weight proteins(3).
  7. α-Lytic Protease is active over a wide temperature range from 4ºC – 50ºC, with optimal activity at 37ºC. Lower temperatures may require longer incubation times.

References

  1. Lumpkin, RJ. et al. (2017). Nat Commun.. 8 (1), 1171.
  2. Meyer, JG. et al. (2014). Molecular & Cellular Proteomics. 13 (3), 823-835.
  3. Ruse, C. New England Biolabs, Inc.. Unpublished observation

Protocols, Manuals & Usage

Protocols

  1. α-Lytic Protease In-gel Digestion Protocol (NEB #P8113)
  2. α-Lytic Protease Typical Reaction Protocol (NEB #P8113)

Tools & Resources

Selection Charts

FAQs & Troubleshooting

FAQs

  1. What reaction buffers are recommended for use with α-Lytic Protease?
  2. Can I perform my digest with α-Lytic Protease at temperatures other than 37°C?
  3. Why should I use a-Lytic Protease instead of Trypsin
  4. Is there an advantage to treating my sample with both α-Lytic Protease and Trypsin?
  5. What reagents inhibit α-Lytic Protease activity?
  6. What reagents stimulate α-Lytic Protease activity?
  7. Can α-Lytic Protease be heat-inactivated?

Quality, Safety & Legal

Quality Assurance Statement

Quality Control tests are performed on each new lot of NEB product to meet the specifications designated for it. Specifications and individual lot data from the tests that are performed for this particular product can be found and downloaded on the Product Specification Sheet, Certificate of Analysis, data card or product manual. Further information regarding NEB product quality can be found here.

Specifications

The Specification sheet is a document that includes the storage temperature, shelf life and the specifications designated for the product. The following file naming structure is used to name these document files: [Product Number]_[Size]_[Version]

Certificate Of Analysis

The Certificate of Analysis (COA) is a signed document that includes the storage temperature, expiration date and quality controls for an individual lot. The following file naming structure is used to name these document files: [Product Number]_[Size]_[Version]_[Lot Number]

Safety DataSheets

The following is a list of Safety Data Sheet (SDS) that apply to this product to help you use it safely.

Legal and Disclaimers

Products and content are covered by one or more patents, trademarks and/or copyrights owned or controlled by New England Biolabs, Inc (NEB). The use of trademark symbols does not necessarily indicate that the name is trademarked in the country where it is being read; it indicates where the content was originally developed. The use of this product may require the buyer to obtain additional third-party intellectual property rights for certain applications. For more information, please email busdev@neb.com.

This product is intended for research purposes only. This product is not intended to be used for therapeutic or diagnostic purposes in humans or animals.

New England Biolabs (NEB) is committed to practicing ethical science – we believe it is our job as researchers to ask the important questions that when answered will help preserve our quality of life and the world that we live in. However, this research should always be done in safe and ethical manner. Learn more.

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