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Endoproteinase AspN |NEB酶试剂 New England Biolabs

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产品信息

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

产品来源

Purified from Flavobacterium menigosepticum.

重组

Endoproteinase AspN should be reconstituted by the addition of 50-500 µl of high purity water. Rapid autolysis is a function of enzyme concentration.

产品类别:
Proteases Products,
Proteome Analysis Products

应用:
Glycomics and glycoproteomics,
Proteomics

  • 产品组分信息

    本产品提供以下试剂或组分：

    NEB # 名称 组分货号 储存温度 数量 浓度
    • 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

  • 特性和用法

    反应条件

    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)

    使用浓度

    100 ng/μl

    分子量

    理论上的: 40089.9 daltons

  • 相关产品

    相关产品

    • Endoproteinase GluC
    • Trypsin-ultra™, Mass Spectrometry Grade
    • Endoproteinase LysC
    • α-Lytic Protease

  • 注意事项

    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).

  • 参考文献

    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.

操作说明、说明书 & 用法

  • 操作说明

    1. Protocol using Endoproteinase AspN (NEB #P8104)

工具 & 资源

  • 选择指南

    • Protease Selection Chart
    • Substrate-based Ligase Selection Chart

FAQs & 问题解决指南

  • 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.