Endoproteinase AspN

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

Companion Products

• Endoproteinase GluC
• Trypsin-ultra™, Mass Spectrometry Grade
• Endoproteinase LysC
• α-Lytic Protease
• Trypsin-digested BSA MS Standard (CAM-modified)

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).

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.

1. Protocol using Endoproteinase AspN (NEB #P8104)

• Protease Selection Chart
• Substrate-based Ligase Selection Chart

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.

• Glycoproteomics Brochure
• Glycoproteomics Technical Guide

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.

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]

• P8104S_v2

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]

• P8104S_v1_10017067
• P8104S_v1_10039459
• P8104S_v1_10047006
• P8104S_v1_10056658
• P8104S_v1_10062862
• P8104S_v1_10070005
• P8104S_v1_10084161
• P8104S_v1_10106640
• P8104S_v1_10118371
• P8104S_v2_10128978
• P8104S_v2_10140339
• P8104S_v2_10158918
• P8104S_v2_10162041
• P8104S_v2_10189623
• P8104S_v2_10215179
• P8104S_v1_0041801
• P8104S_v2_10226516
• P8104S_v2_10226730
• P8104S_v2_10241006
• P8104S_v2_10256174
• P8104S_v2_10269799
• P8104S_v2_10275166
• P8104S_v2_10284689
• P8104S_v2_10298489
• P8104S_v2_10308751
• P8104S_v2_10324292
• P8104S_v2_10332562

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

• Endoproteinase AspN
• Endoproteinase AspN Reaction Buffer

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