标签归档：endoproteinase

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.

Endoproteinase GluC |NEB酶试剂 New England Biolabs

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上海金畔生物科技有限公司代理New England Biolabs（NEB）酶试剂全线产品，欢迎访问官网了解更多产品信息和订购。

产品信息

Endoproteinase GluC (Staphylococcus aureus Protease V8) is a serine proteinase which selectively cleaves peptide bonds C-terminal to glutamic acid residues (1). Endoproteinase GluC also cleaves at aspartic acid residues at a rate 100-300 times slower than at glutamic acid residues (2,3).

Endoproteinase GluC |
Endoproteinase GluC |
A 16 hour GluC digestion of Equine heart cytochrome c using a 1:20 enzyme to substrate ratio, followed by LC-MS with a Thermo™ LTQ Orbitrap XL, identified 97 peptides with 77% sequence coverage.

产品来源

Staphylococcus aureus Protease V8 gene cloned and expressed with histidine-tag in Bacillus subtilis

重组

Endoproteinase GluC should be reconstituted by the addition of 50-500 µl of high purity water. Finger flick the volume of water in the tube to fully resuspend the enzyme. Rapid autolysis is a function of enzyme concentration; any sample reconstituted in a small volume should be used immediately. To get the most use out the enzyme, resuspend the enzyme in 500 μl H2O and aliquot 50 ul each in 10 tubes. Freeze the tubes that are not being used immediately at -20°C for up to two weeks or less. Storage at – 80°C will prolong enzyme stability approximately 2-4 additional weeks.

产品类别:
Proteases Products,
Proteome Analysis Products

应用:
Glycomics and glycoproteomics,
Protein Digestion

  • 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
    • P8100S     -20    
        Endoproteinase GluC P8100SVIAL -20 1 x 50 µg Not Applicable
        GluC Reaction Buffer B8100SVIAL -20 1 x 1.5 ml 2 X

  • 特性和用法

    反应条件

    1X GluC Reaction Buffer
    Incubate at 37°C

    2X GluC Reaction Buffer
    50 mM Tris-HCl
    0.5 mM Glu-Glu
    (pH 8 @ 25°C)

    使用浓度

    100 ng/μl

    分子量

    理论上的: 29849 daltons

  • 相关产品

    相关产品

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

  • 注意事项

    1. Storage conditions: Supplied in dry format from Tris-HCl and sodium chloride buffer.
    2. Repeated freeze/thawing is not recommended.
    3. GluC may also be used in 20-50mM phosphate or ammonium bicarbonate buffer. For optimal activity, supplement reactions with Glu-Glu dipeptide. Glutamic acid residues are highly favored in Tris, phosphate and ammonium bicarbonate buffers.
    4. GluC is purified with a His-tag at its C-terminal of the protein. The predominant species has four histidine residues at the C-terminus with the remaining species showing a distribution from one to six His tags.

  • 参考文献

    1. Drapeau, G.R., Boily, Y. and Houmard, J. (1972). Purification and properties of an extracellular protease of Staphylococcus aureus. J. Biol. Chem.. 247, 6720-26.
    2. Benner, J.S., Martin, D. and Schampel, A.K. unpublished results.
    3. Birktoft, J.J. and Breddam, K. (1994). Proteolytic Enzymes: Serine and Cysteine Peptidases. A.J. Barrett(Ed.), Glutamyl endopeptidases. Methods Enzymol.. 244, 114-126. San Diego: Academic Press.
    4. Breddam, K. and Meldal, M. (1992). Substrate preference of glutamic-acid-specific endopeptidases assessed by synthetic peptide substrates based on intramolecular fluorescence quenching. Eur. J. Biochem.. 206, 103-7.

操作说明、说明书 & 用法

  • 操作说明

    1. Typical GluC Digest Reaction Conditions (P8100)
    2. Typical Trypsin and GluC Co-Digest Reaction Conditions (P8100)
    3. Typical GluC In-Gel Digest Reaction Conditions (P8100)

工具 & 资源

  • 选择指南

    • Protease Selection Chart

FAQs & 问题解决指南

  • FAQs

    1. What is the stability/compatibility of Endoproteinase GluC with regards to presence of DTT, urea, concentrations of methanol, acetonitrile, nitrosoguanidine, thiourea and glycerol?
    2. How does one carry out limited proteolysis experiments with Trypsin and Endoproteinase Glu-C? Which chemicals can be used to quench the reactions at given time points to monitor proteolysis progress via SDS- PAGE?

Endoproteinase LysC |NEB酶试剂 New England Biolabs

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上海金畔生物科技有限公司代理New England Biolabs（NEB）酶试剂全线产品，欢迎访问官网了解更多产品信息和订购。

产品信息

Endoproteinase LysC is a serine endoproteinase, isolated from Lysobacter enzymogenes, that cleaves peptide bonds at the carboxyl side of lysine. LysC is a sequencing grade enzyme and is suitable for proteomics and glycobiology applications.

Endoproteinase LysC |
A 16 hour native LysC digestion of Issatchenkia orientalis Cytochrome c using a 1:50 enzyme to substrate ratio, followed by LC-MS with a Thermo™ LTQ Orbitrap XL, identified 79 peptides with 85% sequence coverage.

产品来源

Isolated from Lysobacter enzymogenes

重组

Endoproteinase LysC should be reconstituted in 200 μl double-distilled water to make a 100 ng/μl solution in 10 mM Tris-HCl, pH 8.0. Rapid autolysis is a function of enzyme concentration.

产品类别:
Proteases Products,
Proteome Analysis Products

应用:
Glycomics and glycoproteomics,
Protein Digestion,
Glycobiology & Proteomics,

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
    • P8109S     -20    
        Endoproteinase LysC P8109SVIAL -20 1 x 0.02 mg Not Applicable

  • 特性和用法

    使用浓度

    100 ng/μl

    分子量

    实际: 30000 daltons

  • 优势和特性

    应用特性

    • Digestion of proteins for proteomic analysis by Mass Spectrometry
    • Protein and peptide identification

  • 相关产品

    相关产品

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

  • 注意事项

    1. Storage Conditions: Supplied in dry format from a Tris-HCl buffer. The solution can be stored at 4°C for several days or in single-use aliquots at –20°C for several months. Use only freshly reconstituted protease for best results.
    2. LysC is active over a wide pH range, pH 5-12, depending on the application. Ammonium bicarbonate buffer may also be used in place of Tris-HCl.

操作说明、说明书 & 用法

  • 操作说明

    1. In-gel Digestion Protocol for Endoproteinase LysC (P8109)
    2. Typical Reaction Conditions for Endoproteinase LysC (P8109)

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. What buffer conditions can Endoproteinase LysC be used in? Are there any special applications for which LysC is especially good?
    2. How much Endoproteinase LysC do I really need to digest my protein?
    3. Can Endoproteinase LysC used in a double digest with Trypsin-ultra?
    4. Can Endoproteinase LysC be used in acidic buffer conditions?

Endoproteinase Glu C, Sequencing grade 测序级谷氨酰蛋白内切酶Glu-C

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Endoproteinase Glu C, Sequencing grade

测序级谷氨酰蛋白内切酶Glu-C

品牌：FUJIFILM Wako
CAS No.：66676-43-5
储存条件：2-10℃
纯度：–
产品编号

(生产商编号)

 等级 规格 运输包装 零售价(RMB) 库存情况 参考值

050-05941

 for Biochemistry 50ug – 4,490.00 – –


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