计算溶液所需的质量、体积或浓度。
为什么选择此级别
，适用于对基线干扰要求严格的色谱和分析工作流程。
储存与运输
2-8°C储存。低温运输 。请查阅批次 COA 获取详细规格。
质量文档
SDS、COA、产品数据表及规格说明书均可下载。可通过批号查询获取批次 COA。
文献证明
在色谱分析、有机合成和交叉偶联反应领域已被 0 篇同行评审文献引用。
概述
Product Description
Endo H, endo-beta-N-acetylglucosaminidase H, Endoglycosidase H
Endo H cleaves Asparagine-linked hybrid or high mannose oligosaccharides, but not complex oligosaccharides. It cleaves between the two N-acetylglucosamine residues in the diacetylchitobiose core of the oligosaccharide, generating a truncated sugar molecule with one N-acetylglucosamine residue remaining on the asparagine. In contrast, PNGase F removes the oligosaccharide intact. Detergent and heat denaturation may increase the rate of cleavage for some glycoproteins.
Recommended Reagents
included with:
1 vial: 5x Reaction Buffer - 400 ml
250 mM sodium phosphate, pH5.5
1 vial: Denaturation Solution - 200 ml 2%
SDS, 1 M Beta-mercaptoethanol
Molecular weight
approximately 29 kD
pH optimum:
5.5, active over the range 5-6.
Applications
Releases asparagine-linked hybrid or high mannose oligosaccharides, but not complex oligosaccharides. Endo H cleaves between the two N-acetylglucosamine residues in the diacetylchitobiose core of the oligosac-charide, generating a truncated sugar molecule with one N-acetylglucosamine residue remaining on the asparagine. In contrast, PNGase F removes the oligosaccharide intact. Detergent and heat denaturation may increase the rate of cleavage for some glycoproteins.
Specific Activity
One unit of Endo H activity is defined as the amount of enzyme required to catalyze the release of N-linked oligosaccharides from 1 µmole of denatured Ribonuclease B in one minute at 37˚C, pH 5.5. Cleavage is monitored by SDS-PAGE (cleaved Ribonuclease B migrates faster).
Specificity
Endo H cleaves Asparagine-linked hybrid or high mannose oligosaccharides, but not complex oligosaccharides. It cleaves between the two N-acetylglucosamine residues in the diacetylchitobiose core of the oligosaccharide, generating a truncated sugar molecule with one N-acetylglucosamine residue remaining on the asparagine. In contrast, PNGase F removes the oligosaccharide intact. Detergent and heat denaturation may increase the rate of cleavage for some glycoproteins.
Formulation
The enzyme is provided as a sterile-filtered solution in 20 mM Tris-HCl, 25mM NaCl, 1 mM EDTA (pH 7.5).
Stability
Several days exposure to ambient temperatures will not reduce activity. Stable at least 12 months when stored properly.
Quality & Purity
Endo H is tested for contaminating protease as follows: 10 µg of denatured BSA is incubated at 37°C for 24 hours with 2 µl of enzyme. SDS-PAGE analysis of the treated BSA shows no evidence of degradation. The production host strain has been extensively tested and does not produce any detectable glycosidases.
Directions for use
1. Add up to 200 µg of glycoprotein to an Eppendorf tube. Adjust to 37.5 µl final volume with de-ionized water.
2. Add 10 µl 5x Reaction Buffer 5.5 and 2.5 µl of Denaturation Solution. Heat at 100°C for 5 minutes.
NOTE: It is not necessary to add Triton X-100. SDS will not inactivate Endo H.
3. Add 2.0 µl of Endo H to the reaction. Incubate 3 hours at 37°C.
If SDS or heat denaturation is omitted, increase incubation time to at least 24 hours.
Monitor cleavage by SDS-PAGE.
The production host strain has been extensively tested and does not produce any detectable glycosidases.
规格
产品名称
Endo-H
别名
内切酶
英文别名
endo-beta-N-acetylglucosaminidase H
规格或纯度
Specific Activity ＞40 U/mg;Activity ＞5 U/ml
生物活性
Activity ≥ 5 U/ml
酶学委员会编号
3.2.1.96
分子类型
小分子
产品线
生物产品
储存与运输
浓度
Specific Activity ＞40 U/mg;Activity ＞5 U/ml
储存条件
2-8°C储存
运输条件
低温运输
技术文档
高级数据
质检证书（CoA,COO,BSE/TSE 和分析图谱)
C of A & Other Certificates(BSE/TSE, COO):
输入批号以搜索分析图谱:
溶液计算器
产品仅提供给经过验证的企业、机构和合格的专业人员用于研发。不适用于人类、动物、诊断或治疗。
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