Trypsin-ultra™, Mass Spectrometry Grade |NEB酶试剂 New England Biolabs

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产品信息

Trypsin-ultra™, Mass Spectrometry Grade is a serine endopeptidase. It selectively cleaves peptide bonds C-terminal to lysine and arginine residues (1). Trypsin-ultra is treated with L-(tosylamido-2-phenyl) ethyl chloromethyl ketone (TPCK) to inactivate any remaining chymotryptic activity. It is modified by acetylation of the ε-amino groups of lysine residues to prevent autolysis. Trypsin-ultra (TPCKtreated) cleaves at Lys-Pro and Arg-Pro bonds at a much slower rate than other amino acid residues (2).

Digestion with Trypsin-ultra results in a high number of proteins identified, and sequence coverages as high as 90%

Trypsin-ultra™, Mass Spectrometry Grade |

A 3-hour Trypsin-ultra digest of Pyrococcus furiosus using a 1:50 enzyme:substrate ratio, followed by ESI-MS, resulted in 418 proteins identified with individual protein sequence coverage as high as 90%.

产品来源

Isolated from bovine (Bos taurus) pancreas.

重组

Trypsin-ultra, Mass Spectrometry Grade should be reconstituted by the addition of 20–200 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,
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
    • P8101S     -20    
        Trypsin-ultra™, Mass Spectrometry Grade P8101SVIAL -20 5 x 20 µg Not Applicable
        Trypsin-ultra, Reaction Buffer B8101SVIAL -20 1 x 1.5 ml 2 X

  • 特性和用法

    反应条件

    1X Trypsin-ultra, Reaction Buffer
    Incubate at 37°C

    1X Trypsin-ultra, Reaction Buffer
    50 mM Tris-HCl
    20 mM CaCl2
    (pH 8 @ 25°C)

    使用浓度

    100 ng/μl

    分子量

    理论上的: 23675 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

    • IdeZ Protease (IgG-specific)
    • p6043-rapid-pngase-f-antibody-standard
    • Endo S
    • Rapid 快速 PNGase F
    • p0711-rapid-pngase-f-non-reducing-format
    • Endoproteinase GluC
    • Endoproteinase AspN
    • Endoproteinase LysC
    • α-Lytic Protease

  • 注意事项

    1. Substrate must be in phosphate-free buffer to prevent calcium precipitation with both reconstituted enzyme and enzyme buffer.
    2. Trypsin-ultra, Mass Spectrometry Grade is acetylated on multiple lysine residues. This protein appears as a single band on SDS-PAGE. This sequence is also available at www.neb.com.
    3. Storage Conditions: Supplied in dry format from a sodium acetate and calcium chloride buffer. Store at -20°C.
    4. 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.

  • 参考文献

    1. Northrop, J.H. and Kunitz, M. (1931). Isolation of protein crystals possessing tryptic activity. Science. 73,
    2. Perona, J.J. and Craik, C.S. (1995). Structural basis of substrate specificity in the serine proteases. Protein Sci.. 4, 337-360.

操作说明、说明书 & 用法

  • 操作说明

    1. Protocol using Trypsin-ultra™, Mass Spectrometry Grade (P8101)
    2. Trypsin Digestion Protocol using NEB Trypsin-ultra™ and the FASP Kit

  • 应用实例

    • AppNote_Unbiased_and_Fast_IgG_Deglycosylation _for_Accurate_N-glycan_Analysis_using_Rapid_PNGase_F
    • AppNote_Glycan_Analysis_of_Murine_IgG2a_by_Enzymatic_Digestion_with_PNGase_F_and_Trypsin
    • Glycan Analysis of Murine IgG2a by Enzymatic Digestion with PNGase F and Trypsin, Followed by Mass Spectrometric Analysis

工具 & 资源

  • 选择指南

    • Protease Selection Chart

FAQs & 问题解决指南

  • FAQs

    1. Is there a compatible buffer for digesting with Trypsin (P8101S) and Endoproteinase GluC (P8100S) together?
    2. I have a very low concentration of protein and would prefer not to denature as a separate step with buffer exchange before digestion. What denaturants can he use in the Trypsin reaction itself?
    3. I would like to use Trypsin to digest some proteins which are present in a cell culture supernatant. The supernatant is composed of DMEM supplemented with 0.2% BSA and antibiotics. Will the enzyme work in the cellular supernatant? The final reaction volume will be 100-300 ul.
    4. I am using Trypsin and am wondering about specificity. Does it cut at additional sites when in high concentration?
    5. I want to know if calcium can be left out of the buffer since it is causing my DNA-protein complex to precipitate.
    6. I want to use Trypsin on 20 ug of protein. How much enzyme do I need to use?
    7. How does one do a Trypsin in-gel digest?
    8. Is there a simple way to remove Trypsin after protein cleavage?