Product Class: Other

Factor Xa Protease

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Product Introduction

Factor Xa cleaves after the arginine residue in its preferred cleavage site Ile-Glu/Asp-Gly-Arg. It will sometimes cleave at other basic residues, depending on the conformation of the protein substrate. The most common secondary site, among those that have been sequenced, is Gly-Arg.

Catalog # Size Concentration
P8010S 50 µg 1 mg/ml
P8010L 250 µg 1 mg/ml

Product Information

Description

Factor Xa cleaves after the arginine residue in its preferred cleavage site Ile-(Glu or Asp)-Gly-Arg. It will sometimes cleave at other basic residues, depending on the conformation of the protein substrate (1,2,3). The most common secondary site, among those that have been sequenced, is Gly-Arg. There seems to be a correlation between proteins that are unstable in E.coli and those that are cleaved by Factor Xa at secondary sites; this may indicate that these proteins are in a partially unfolded state (Walker, I., Riggs, P., unpublished observations). Factor Xa will not cleave a site followed by proline or arginine.

Product Source

Factor Xa Protease is purified from bovine plasma and activated by treatment with the activating enzyme from Russell's viper venom.
This product is related to the following categories:
Proteases,
This product can be used in the following applications:
Target Protein Insolubility ,
Fusion Protein Cleavage,
Protein Purification,
Protein Digestion,
Protein Expression

Reagents Supplied

Reagents Supplied

The following reagents are supplied with this product:

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

Properties & Usage

Unit Definition

1 μg of Factor Xa will cleave 50 μg of MBP-fusion protein test substrate, MBP-FXa-Paramyosin-ΔSal to 95% completion in a total reaction volume of 50 μl in 6 hours or less at 23°C in 20 mM Tris-HCl (pH 8.0 @ 25°C) with 100 mM NaCl and 2 mM CaCl2.

Usage Concentration

1 mg/ml

Removal

Factor Xa will bind specifically to benzamidine-agarose.

Storage Buffer

50 mM HEPES
500 mM NaCl
2 mM CaCl2
50% Glycerol
pH 8 @ 25°C

Molecular Weight

Theoretical: 43 kDa

Deactivation

Dansyl-glu-gly-arg-chloromethyl ketone (CALBIOCHEM, #251700) will irreversibly inactivate Factor Xa by covalent attachment at the active site. In a reaction containing 20 μg/ml Factor Xa, 2 μM dansyl-glu-gly-arg-chloromethyl ketone will inactivate >95% of the Factor Xa in 1 minute at room temperature.

Product Notes

  1. The test substrate MBP-ΔSal is maltose-binding protein fused to a truncated form of paramyosin, with the amino acids Ile-Glu-Gly-Arg at the fusion joint. Greater than 95% of the fusion protein is cleaved in 6 hours or less.

References

  1. Nagai, K. et al. (1985). Proc. Natl. Acad. Sci. USA. 82(21), 7252-5.
  2. Quinlan, R.A. et al (1989). J. Cell. Sci. 93 (Pt 1). 71-83,
  3. Eaton, D. et al (1986). Biochemistry. 25, 505-512.

Protocols, Manuals & Usage

Protocols

  1. Cleavage of the Fusion Protein Generated Using The pMAL Protein Fusion and Purification System (E8200)

Tools & Resources

Selection Charts

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.