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Endo H

Endo H

货号: UA070040
价格: 350
规格: 10KU
介绍: -
其他: -
产品规格
  • 物种

    Streptomyces picatus
  • 分子别名

    Endo-beta-N-acetylglucosaminidase H, Endoglycosidase H, Endo H
  • 表达宿主

    E.coli
  • 分子量

    30kD (Reducing)

  • 纯度

    >95% by SDS-PAGE and HPLC
  • 活性

    500U/μl
  • 标记

    Unconjugated
  • 标签

    His Tag
  • 性状

    Liquid
  • 缓冲体系

    20 mM Tris-HCl、50 mM NaCl、5 mM EDTA(pH 7.5 @ 25°C)
  • 储存条件

    Store at -25 ~ -15℃ for 2 years

  • 文献引用

    [1] Wang F , Wang X , Yu X ,et al. High-Level Expression of Endo-β-N-Acetylglucosaminidase H from Streptomyces plicatus in Pichia pastoris and Its Application for the Deglycosylation of Glycoproteins[J].Plos One, 2015, 10.

    [2] Maley F, Trimble R B, Tarentino A L,et al. Characterization of glycoproteins and their associated oligosaccharides through the use of endoglycosidases.[J]. Analytical Biochemistry,1989,180(2):195-204.

  • 稀释度

背景介绍
  • Endo H is a recombinant glycosidase cloned from Streptomyces plicatus and overexpressed in E.coli. It cleaves the chitobiose core of high-mannose oligosaccharides and a limited number of hybrid oligosaccharides from asparagine-linked glycoproteins, but not complex, oligosaccharides from glycoproteins.

产品组分
  • Storage Solution: 500U/μL EndoH、20 mM Tris-HCl、50 mM NaCl、5 mM EDTA (pH 7.5@ 25°C)

    10*Denaturing Buffer: 5% SDS、400 mM DTT

    10*Reaction Buffer: 500 mM sodium acetate(pH 6 @ 25°C)

操作步骤
  • 1. Combine 1-20 μg of glycoprotein, 1 μl of 10*Denaturing Buffer and H20 (if necessary) to make a 10 μl total reaction volume.

    2. Denature glycoprotein by heating raection at 100°C for 10 minutes.

    3. Make a total reaction volume of 20 μl by adding 2 μl of 10*Reaction Buffer, H20 and 1-5 μl Endo H.

    4. Incubate reaction at 37°C for 1 hour.

注意事项
  • Please avoid repeated freeze-thaw cycles.

酶活定义
  • One unit is defined as the amount of enzyme required to remove > 95% of the carbohydrate from 10µg of denatured RNase B in 1 hour at 37°C in a total reaction volume of 10µl.
  • 生物活性JSON

    • In the experimental design, 125 U, 25 U, 5 U, 1 U and 0.2 U enzymes were added to the 20 ul reaction system to investigate the effect of Endo H enzyme on the enzymatic digestion of substrate RNase B.

      M marker

      Lan1 blank

      Lan2 Rnase B 10μg

      Lan3 Rnase B 10μg +125U Endo H

      Lan4 Rnase B 10μg+ 25U Endo H

      Lan5 Rnase B 10μg+ 5U Endo H

      Lan6 Rnase B 10μg+ 1U Endo H

      Lan7 Rnase B 10μg+ 0.2U Endo H

  • 电泳JSON

  • 体积排阻色谱JSON(SEC-HPLC)