Product Class: Other

Endoproteinase AspN
NEBU cloned at NEB 37

P8104_cutsite

Product Introduction

Endoproteinase AspN (flavastacin) is a zinc metalloendopeptidase which selectively cleaves protein and peptide bonds N-terminal to aspartic acid residues.

  • Free of glycerol and detergents which may interfere with mass spectrometry or liquid chromatography methods.
  • Supplied in dry format
  • Optimal activity and stability for up to 12 months
Catalog # Size Concentration
P8104S 50.0 µg

Product Information

Description

Endoproteinase AspN (flavastacin) is a zinc metalloendopeptidase which selectively cleaves peptide bonds N-terminal to aspartic acid residues(1).

Product Source

Purified from Flavobacterium menigosepticum.

Reconstitution


Endoproteinase AspN should be reconstituted by the addition of 50-500 µl of high purity water. Rapid autolysis is a function of enzyme concentration.
This product is related to the following categories:
Proteases Products,
Proteome Analysis Products,
This product can be used in the following applications:
Glycomics and glycoproteomics,
Proteomics

Reagents Supplied

Reagents Supplied

The following reagents are supplied with this product:

NEB # Component Name Component # Stored at (°C) Amount Concentration
  • P8104S     -20    
      Endoproteinase AspN P8104SVIAL -20 1 x 50 µg Not Applicable
      Endoproteinase AspN Reaction Buffer B8104SVIAL -20 1 x 1.5 ml 2 X

Properties & Usage

Reaction Conditions

1X Endoproteinase AspN Reaction Buffer
Incubate at 37°C

1X Endoproteinase AspN Reaction Buffer
50 mM Tris-HCl
2.5 mM ZnSO4
(pH 8 @ 25°C)

Usage Concentration

100 ng/μl

Molecular Weight

Theoretical: 40089.9 daltons

Product Notes

  1. Storage Conditions: Supplied in dry format. Can be stored frozen in solution at -20°C for up to 2 weeks. A decrease in activity will occur if stored in solution. Use only freshly reconstituted protease for best results.
  2. Endoproteinase AspN contains a O-linked carbohydrate on the protein. The protein appears as a single band by SDS-PAGE analysis.
  3. Aspartic acid residues are strongly favored by Endoproteinase AspN in all buffer conditions we have examined (Tris-HCl, ammonium bicarbonate and potassium phosphate) (2).

References

  1. Tarentino A.L. Flavastacin (2004). In Handbook of Proteolytic Enzymes. 2nd Ed., 631-632, Elsevier, London.
  2. Tarentino, A.L. et al. (1995). Arch biochem biophys . 319, 281-285.
  3. Grimwood, B.G., Plummer, T.H. and Tarentino, A.L. (1994). Arch Biochem Biophys . 311, 127-132.

Protocols, Manuals & Usage

Protocols

  1. Protocol using Endoproteinase AspN (NEB #P8104)

Tools & Resources

Selection Charts

FAQs & Troubleshooting

FAQs

  1. Is your Endoproteinase AspN the same as the Endoproteinase AspN sold by other companies?
  2. The Endoproteinase AspN is not fully dissolving when I reconstitute it in water. How do I get it completely into solution?
  3. Which residues does Endoproteinase AspN cut?
  4. Do you have a protocol or pointers for digesting proteins in-gel with endo- proteinase Asp-N followed by Endoproteinase Glu-C? I'm following the protocol for each but would like to streamline the combined digest.
  5. How pure is Endoproteinase AspN? Has it been checked for nuclease activity?
  6. Tris will interfere with my reactions downstream. Does your Endoproteinase AspN, when reconstituted in H2O, contain Tris-HCl?
  7. What is the Endoproteinase AspN sequence in text format?
  8. Can Endoproteinase AspN be used with a volatile buffer?
  9. How stable is this enzyme? We dissolved our enzyme in water approx 2 weeks ago and the literature indicates that activity will start to drop after this length of storage time(and longer).
  10. I digested BSA with Endoproteinase AspN following standard conditions and examined the released fragments with Mass-spec. I did not see many peaks that matched the predicted fragment but saw a lot of unpredicted bands, perhaps indicating non-specific enzyme activity.
  11. Can endoproteinase AspN be heat inactivated?
  12. I can’t get Endoproteinase AspN to digest my protein.

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 [email protected].

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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