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Trypsin, Mass Spectrometry Grade

Trypsin(pig),porcine pancreas,Trypsin,Mass Spectrometry Grade

价格 ￥280.00 （ 供应商现货 ： 3-5个工作日）
货号 UA070130
规格
数量
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产品介绍

产品介绍 评论(0)

产品规格

  • 分子别名

    Trypsin(pig); porcine pancreas; Trypsin，Mass Spectrometry Grade

  • 分子量

    24±2 kDa (Reducing)

  • 纯度

    ＞95% by SDS-PAGE

  • 活性

    ≥6800 USP/mg

  • 标签

    No Tag

  • 性状

    Lyophilized Powder

  • 缓冲体系

    2 mM HCl, 2 mM CaCl2

  • 溶解方法

    Trypsin, Mass Spectrometry Grade should be dissolved in 2 mM HCl to prepare a 1 mg/mL solution for subsequent use.

  • 储存条件

    Store at -25 ~ -15℃ for 2 years

  • 文献引用

    Olsen, J. V. Trypsin Cleaves Exclusively C-terminal to Arginine and Lysine Residues[J]. Molecular & Cellular Proteomics, 2004, 3(6):608-614.
    Chi W J. Medium Optimization and Application of Affinity Column Chromatography for Trypsin Production from Recombinant Streptomyces griseus[J]. Journal of Microbiology & Biotechnology, 2009, 19(10):1191-1196.

背景介绍

  • Trypsin is a serine protease commonly found in the vertebrate digestive system that hydrolyzes proteins. Active trypsin specifically hydrolyzes peptide bonds on the carboxyl side of lysine or arginine residues, provided the next residue is not proline. This specificity makes trypsin indispensable for the initial digestion of dietary proteins into smaller peptides, which are then further degraded by other proteases. Trypsin consists of two subunits, the alpha subunit (with 2 polypeptide chains forming it) and the beta subunit (with 1 polypeptide chain forming it). Trypsin, Mass Spectrometry Grade was subjected to reductive methylation, producing a highly active, stable molecule that strongly prevented its autolytic activity and reduced interference from excess peptide fragments generated by self-hydrolysis. It can be widely applied in laboratory proteomics analyses such as protein and peptide sequencing identification and HPLC peptide-map analysis.

产品组分

  • Trypsin，Mass Spectrometry Grade lyophilized
    2*Reaction buffer:，100 mM Tris-HCl, 40 mM CaCl2, pH 8.0 @ 25°C

操作步骤

  • 1 Trypsin, Mass Spectrometry Grade should be reconstituted by the addition of 20–200 μl of 2 mM HCl
    2 Set-up a typical reaction as follows
    1）Add the following components in sequence

    Components

    Volume

    Substrate protein

    5 μL (about 2 ug)

    ddH2O

    4.5 μL

    2*Reaction buffer

    10 μL

    100 ng/μL Tyrpsin

    0.5 μL

    2）Incubate at 37°C for 2~4 h

注意事项

  • 1. Substrate must be in phosphate-free buffer to prevent calcium precipitation with both reconstituted enzyme and enzyme buffer.

    2. Storage buffer: 2 mM HCl, 2 mM CaCl2.

酶活定义
  • In a 3.2 mL reaction system using BAEE as substrate, the UV absorption at 253 nm increased by 0.003 per min using cuvette with 1 cm optical path, which was defined as 1 USP trypsin activity unit

  • 生物活性

    • In terms of sequence coverage, UA070130 demonstrates a slight advantage, with enzyme P and enzyme Y exhibiting coverage values that are 1.3% and 2.3% lower, respectively. However, when considering peptides with no more than three missed cleavage sites, a different trend emerges. Enzyme Y yields the highest proportion, followed by enzymes UA070130 and P, which show marginally lower values. These results indicate that the three enzymes display comparable efficacy in protein digestion.

  • 电泳

    • 2μg (R: reducing condition, N: non-reducing condition).

  • 反相高效液相色谱(RP-HPLC)

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