计算溶液所需的质量、体积或浓度。
Recombinant PNGase F
有货
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
Recombinant PNGase F is isolated from a E. coli strain containing a clone of the Elizabethkingia miricola gene. There is no detectable difference in activity or specific activity of the recombinant enzyme from the native enzyme.
PNGase F cleaves asparagine-linked (N-linked) oligosaccharides from glycoproteins. PNGase F deaminates asparagine to aspartic acid, leaving the oligosaccharides intact. Denaturation increases the rate of cleavage up to 100x. Most native proteins can still be completely N-deglycosylated but incubation time must be increased. PNGase F will remain active under incubation conditions for at least 72 hours. PNGase F will not remove oligosaccharides containing Alpha-(1,3)-linked core fucose commonly found on plant glycoproteins; for this purpose, use peptide N-glycosidase A.
Contents
60 µl aliquot of recombinant PNGase F (0.3 U) in 20 mM Tris-HCl, pH 7.5
Included with 20 µL and 60 µL pack sizes
5x PNGase F Reaction Buffer 7.5 for PNGase F – 250 mM sodium phosphate, pH 7.5
PNGase F Denaturation Solution – 2% SDS, 1 M Beta-mercaptoethanol
PNGase F Triton X-100 – 15% solution
Formulation
The enzyme is provided as a sterile-filtered solution in 20 mM Tris-HCl (pH 7.5).
Molecular Weigh
approximately 35 kD.
pH optimum
7.5, active over the range 6-10.
Specific Activity
Specific Activity Defined as the amount of enzyme required to catalyze the release of N-linked oligosaccharides from 1 micromole of RNase B in 1 minute at 37˚C, pH 7.5. Cleavage is monitored by SDS-PAGE (cleaved RNase B migrates faster).
Specificity
PNGase F cleaves asparagine-linked (N-linked) oligosaccharides from glycoproteins. PNGase F deaminates asparagine to aspartic acid, leaving the oligosaccharides intact. Denaturation increases the rate of cleavage up to 100x. Most native proteins can still be completely N-deglycosylated but incubation time must be increased. PNGase F will remain active under incubation conditions for at least 72 hours. PNGase F will not remove oligosaccharides containing Alpha-(1,3)-linked core fucose commonly found on plant glycoproteins; for this purpose, use peptide N-glycosidase A.
Stability
Several days exposure to ambient temperatures will not reduce activity. Stable at least 12 months when stored properly.
Quality & Purity
PNGase F 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.
PNGase F Suggested usage
1. Add up to 200 µg of glycoprotein to an Eppendorf tube. Adjust to 35 µl final volume with de-ionized water.
2. Add 10 µl 5x PNGase F Reaction Buffer 7.5 and 2.5 µl of PNGase F Denaturation Solution. Heat at 100˚C for 5 minutes.
3. Cool. Add 2.5 µl of PNGase F Triton X-100 and mix.
4. Add 2.0 µl of PNGase F 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.
规格
产品名称
Recombinant PNGase F
英文别名
Peptide-N4-(acetyl-ß-glucosaminyl)-asparagine amidase, N-Glycosidase F
规格或纯度
Specific Activity ＞25 U/mg;Activity 5 U/ml
生物活性
5 U/ml
酶学委员会编号
3.5.1.52
分子类型
蛋白质
产品线
生物产品
储存与运输
浓度
Specific Activity ＞25 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:
技术文档和文章
溶液计算器
常见问题
本产品应如何储存？
请于2–8 °C条件下保存。需冷藏保存以维持规定的有效期。
本产品如何运输？
本产品采用湿冰冷藏运输。收货后请立即拆包，并按上述储存条件保存。
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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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